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Bioorganic & Medicinal Chemistry: Christoph Nitsche, Christian Steuer, Christian D. Klein
Bioorganic & Medicinal Chemistry: Christoph Nitsche, Christian Steuer, Christian D. Klein
a r t i c l e i n f o a b s t r a c t
Article history: The 3-aryl-2-cyanoacrylamide scaffold was designed as core pharmacophore for inhibitors of the Dengue
Received 20 September 2011 and West Nile virus serine proteases (NS2B-NS3). A total of 86 analogs was prepared to study the struc-
Revised 18 October 2011 ture–activity relationships in detail. Thereby, it turned out that the electron density of the aryl moiety
Accepted 19 October 2011
and the central double bond have a crucial influence on the activity of the compounds, whereas the influ-
Available online 25 October 2011
ence of substituents of the amide residue is less relevant. The para-hydroxy substituted analog was found
to be the most potent inhibitor in this series with a Ki-value of 35.7 lM at the Dengue and 44.6 lM at the
Keywords:
West Nile virus protease. The aprotinin competition assay demonstrates a direct interaction of the inhib-
Dengue virus
West Nile virus
itor molecule with active centre of the Dengue virus protease. The target selectivity was studied in a
Flavivirin counterscreen with thrombin and found to be 2.8:1 in favor of DEN protease and 2.3:1 in favor of
NS2B-NS3 protease WNV protease, respectively.
Nitrile Ó 2011 Elsevier Ltd. All rights reserved.
Aldol condensation
0968-0896/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2011.10.061
C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337 7319
The compounds were screened against the DEN and WNV pro-
teases (NS2B-NS3) and thrombin (as a counterscreen for selectivity
studies). The results at the different targets are shown in Table 1.
The best activities against DEN and WNV proteases were observed
for the para-hydroxy derivatives 21 and 23–27. Some of those
derivatives show inhibition values of more than 50% at 50 lM
inhibitor and substrate concentration at DEN and WNV proteases.
In general it can be observed that an aromatic system with an elec-
Figure 1. Pharmacophoric model and possible molecular interactions. R1 = aryl, tron withdrawing group resulted in low activity, whereas electron
R2 = H or alkyl, R3 = amine, S1 and S01 describe enzyme pockets, Ser135 is the serine
residue of the catalytic triad.
donor groups like hydroxy or amine substituents gave best results.
Para-substituted aromatic systems showed better activity than
meta-substituted and para,meta-disubstituted systems. There is a
2. Results and discussion loss of nearly 40% of activity if the hydroxy substituent is posi-
tioned in meta (28) and not in para position (21). Various halogen,
2.1. Synthesis amine or methoxy substituents in the para-position were unable to
maintain the activity of the hydroxy-substituted analogs. However,
For the synthesis of 3-aryl-2-cyanoacrylamides a one- or two- non-functionalized aliphatic chains like methyl (5) or isopropyl
step approach was used as shown in Scheme 1. If unsubstituted (6–9) gave satisfying results. Various heterocycles, also shown in
2-cyanoacetamide was used (R3 = NH2), step (b) in Scheme 1 was Table 1, did not result in increased activity. In contrast to the
employed to obtain 3-aryl-2-cyanoacrylamide derivatives. Other- importance of the aryl moiety we only found a limited influence
wise the N-substituted 2-cyanoacetamide had to be synthesized of the amide residue (Table 1). Attempts to mimic the substrate
in a previous reaction step from methyl 2-cyanoacetate in an effi- more closely by attaching a serine residue (for S01 pocket interac-
cient direct treatment with the required amine without solvent at tion) in compounds 16 and 17 were unsuccessful. Apparently, the
room temperature (step (a) in Scheme 1).9 In some cases the S1 and S2 pockets are most important for molecular recognition at
carbonyl component (mostly aromatic aldehydes) had to be syn- the DEN and WNV proteases. We assume that the aryl moiety
thesized from other, commercially available compounds. The aldol interacts with the S1 pocket of the enzyme, probably combined
condensation with aromatic aldehydes was carried out in metha- with an interaction of the electrophilic nitrile and the adjacent cat-
nol with N-methylpiperazine as catalyst leading in a mostly alytic serine (Fig. 1).
selective and quantitative fashion to the trans-olefin product. The An enlargement of inhibitor molecules towards the S2 pocket
trans-geometry of the products was verified by the crystal struc- can be expected to increase the activity and selectivity of the com-
ture of compound 14.10 Only in a few cases a small amount of pounds. Therefore, we synthesized molecules which are able to
cis-olefin was obtained (detected by LC–MS). However, the use of create additional interactions in the direction of the S2 pocket. An
ketones instead of aldehydes gave mixtures of cis and trans isomers extension of the compounds without modification of the underly-
(Table 1). By X-ray crystal structural analysis of compound 3b10 ing arylacrylonitrile pharmacophore is only possible at the olefinic
and NMR correlation, we were able to determine the absolute R2 position or at the aromatic system. Non-hydrogen substituents
structure of all compounds. The approach towards the cyclic N- at R2 caused a pronounced decrease in activity. An additional
arylamines 55–63, 70 and 71 depends on the desired N-aryl- methyl group at R2 (as in compound 22a) leads to a decrease of
connection in ortho, meta or para position. The para-substituted activity of more than 30% in comparison to 21 at all targets. The
compounds 55–60 are available via a one pot/three component geometry of the double bond is apparently not critical in all tested
reaction of the corresponding cyanoacetamide, the 4-fluorobenzal- R1- and R2-disubstituted compounds (2a/b, 3a/b, 4a/b, 22a/b).
dehyde and the secondary cyclic amine as shown in Scheme 2.11 Various extensions of the aromatic system were also evaluated.
We were also able to transfer this reaction to 2-fluorobenzalde- The para-benzyloxy substituted derivative 72 was an initial test for
hyde to obtain the ortho-substituted derivative 63. Pentafluoro- a possible occupation of the S2 pocket. This attempt resulted in an
benzaldehyde as starting material for derivative 61 reacted inactive molecule at both tested viral targets in comparison to its
selectively in para-position at room temperature. The meta-substi- analog 32. Next we evaluated cyclic systems with restricted flexi-
tuted derivative 62 and the heterocyclic compounds 70 and 71 bility. For this we synthesized substituted cyclic N-arylamines
were available by a copper catalyzed coupling reaction of the cyclic (55–63).
amine with the corresponding bromoarylaldehyde.12 The evaluation of the cyclic N-arylamine derivatives showed, in
The aromatic amide derivatives 64–66 were available by amide accordance with the results described before, that para-substituted
coupling reaction of the desired amine and the aromatic carboxylic aromatic systems gave best activity results, whereas ortho- or meta-
acids 37–39. substitution resulted in decreased activities. Best results showed
The double bond of compounds 40, 20, 57, 54, 69 and 67 could compounds 59 and 60 with good activities at the DEN and WNV pro-
be reduced selectively with sodium borohydride in methanol to tease. The perfluorinated compound 61 shows a decline in activity in
obtain the saturated analogs 76–81. comparison to the non-fluorinated derivatives 59 and 60. This is also
in correlation with the results described before for the simple substi-
tuted aryl moieties. In general, it therefore seems obvious that an
aromatic system with high electron density is needed for an effective
interaction with the target. The 4-hydroxy substituted derivatives
remained the most active compounds in this series. Different aro-
matic amide derivatives 64–66 were not able to increase activity.
The same unsatisfying results were observed with the less drug-like
Scheme 1. Synthetic approach for simple 3-aryl-2-cyanoacrylamides. Reagents and
double-unsaturated compounds 67–69.
conditions: (a) no solvent, rt, 20 h; (b) N-methylpiperazine, MeOH, rt, 20 h; (c) Because of the apparent structural analogy of the explored com-
NH4OAc/AcOH, toluene, reflux, 2–5 h. pounds with the known catecholamine O-methyl transferase
Table 1
7320
Activity of substituted arylacrylamides against the Dengue and West Nile virus proteases in comparison to thrombin.
1 H NH2 15.0 ± 1.6 9.3 ± 4.9 12.5 ± 5.2 35 H NH2 n.i. 10.5 ± 2.2 n.i.
2a Me NH2 n.i. 13.7 ± 1.1 7.3 ± 3.9 36 H NH2 13.3 ± 1.9 20.0 ± 6.3 10.6 ± 5.1
3a i-Pr NH2 n.i. n.i. 16.7 ± 9.7 38 H 20.0 ± 1.2 31.0 ± 1.1 15.9 ± 1.5
3b i-Pr NH2 n.i. 13.8 ± 4.6 10.1 ± 2.9 39 H 20.2 ± 2.2 20.1 ± 5.2 11.9 ± 2.4
4a Pr NH2 6.7 ± 1.8 15.3 ± 4.0 11.1 ± 1.1 40 H NH2 35.1 ± 2.4 46.0 ± 0.9 21.6 ± 0.7
4b Pr NH2 7.6 ± 2.7 16.0 ± 1.7 6.9 ± 0.9 41 H 22.6 ± 0.8 33.2 ± 3.1 23.7 ± 2.5
5 H NH2 32.2 ± 5.4 22.8 ± 3.7 n.i. 42 H 23.3 ± 3.8 28.1 ± 3.7 19.8 ± 3.5
6 H NH2 23.0 ± 3.3 24.6 ± 3.3 19.4 ± 5.2 43 H 25.3 ± 4.5 21.3 ± 3.5 n.i.
7 H 18.2 ± 4.5 22.2 ± 4.2 17.9 ± 5.4 44 H 43.3 ± 2.7 38.1 ± 2.0 25.1 ± 2.9
8 H 28.2 ± 1.2 46.3 ± 1.0 14.4 ± 6.0 45 NH2 23.2 ± 3.1 24.1 ± 2.5 10.3 ± 2.1
9 H 24.9 ± 1.3 39.5 ± 1.7 11.6 ± 4.1 46 H NH2 n.i. n.i. n.i.
10 H NH2 22.1 ± 2.4 21.7 ± 0.8 8.7 ± 1.2 47 NH2 n.i. n.i. n.i.
Table 1 (continued)
11 H NH2 22.5 ± 7.4 16.0 ± 2.1 7.9 ± 5.5 48 H NH2 28.4 ± 7.9 23.9 ± 2.5 15.7 ± 3.7
12 H NH2 14.7 ± 2.3 10.7 ± 3.0 8.5 ± 2.1 49 H NH2 22.0 ± 7.3 23.8 ± 3.8 27.9 ± 1.3
13 H NH2 11.6 ± 2.0 15.9 ± 2.0 13.6 ± 8.2 50 H NH2 20.2 ± 2.9 19.2 ± 0.8 26.3 ± 0.8
14 H 10.9 ± 0.6 22.6 ± 2.1 n.i. 51 H n.i. 10.5 ± 2.1 17.9 ± 2.2
16 H 20.3 ± 1.3 21.1 ± 1.5 21.4 ± 4.7 53 H 15.6 ± 3.7 23.9 ± 3.7 26.9 ± 0.7
17 H 23.0 ± 1.7 31.7 ± 2.8 23.3 ± 6.0 54 H NH2 23.7 ± 0.6 33.4 ± 2.8 16.4 ± 2.0
18 H NH2 16.3 ± 5.8 12.3 ± 1.7 9.6 ± 7.6 55 H NH2 24.3 ± 8.7 24.6 ± 2.1 17.7 ± 5.8
19 H NH2 19.2 ± 2.7 22.0 ± 2.2 20.0 ± 3.3 56 H NH2 14.0 ± 4.1 24.2 ± 5.4 20.0 ± 7.2
20 H NH2 15.6 ± 2.9 19.3 ± 2.5 17.8 ± 6.5 57 H NH2 28.2 ± 5.8 35.3 ± 3.5 19.2 ± 3.1
21 H NH2 51.7 ± 1.0 55.0 ± 4.6 24.4 ± 4.9 58 H NH2 28.6 ± 3.9 45.0 ± 1.8 26.1 ± 6.5
22a Me NH2 16.8 ± 1.4 21.9 ± 5.7 n.i. 59 H NH2 32.3 ± 1.1 40.9 ± 2.4 22.0 ± 3.0
22b Me NH2 13.2 ± 1.6 22.7 ± 1.9 n.i 60 H 37.4 ± 1.1 47.8 ± 2.5 21.8 ± 3.0
7321
7322
Table 1 (continued)
23 H 44.7 ± 0.7 57.3 ± 5.8 18.8 ± 1.7 61 H NH2 19.2 ± 2.6 31.9 ± 2.7 28.5 ± 3.8
24 H 46.4 ± 0.1 44.4 ± 1.0 16.8 ± 0.6 62 H 18.5 ± 4.8 20.5 ± 1.7 9.7 ± 4.6
25 H 46.6 ± 4.9 41.8 ± 1.7 18.9 ± 3.5 63 H NH2 10.4 ± 3.0 7.7 ± 2.1 7.4 ± 0.3
26 H 32.2 ± 7.9 49.9 ± 4.9 18.0 ± 1.9 64 H NH2 22.3 ± 8.5 24.0 ± 6.0 n.i.
28 H NH2 15.7 ± 1.8 18.1 ± 2.4 9.8 ± 9.7 66 H 10.8 ± 1.8 19.1 ± 1.8 8.0 ± 2.9
29 H NH2 34.9 ± 2.0 28.8 ± 1.7 18.4 ± 9.7 67 H NH2 30.2 ± 1.1 34.6 ± 0.8 20.5 ± 1.3
30 H NH2 24.3 ± 1.7 28.2 ± 0.4 22.5 ± 2.0 68 H 36.8 ± 3.4 30.1 ± 2.6 28.7 ± 2.1
31 H 21.0 ± 1.7 32.4 ± 0.7 21.9 ± 6.2 69 H 10.3 ± 0.9 17.1 ± 1.3 16.2 ± 7.8
32 H NH2 43.2 ± 6.3 42.9 ± 3.6 17.4 ± 4.7 70 H NH2 23.2 ± 3.8 29.1 ± 1.4 38.6 ± 8.1
33 H NH2 30.5 ± 3.1 43.8 ± 3.7 23.0 ± 3.7 71 H 10.5 ± 5.0 24.0 ± 0.5 47.0 ± 4.0
34 H NH2 43.1 ± 0.1 28.8 ± 3.9 24.4 ± 6.7 72 H NH2 n.i. n.i. 16.0 ± 2.4
a
% Inhibition of the Dengue Virus (DEN) NS2B-NS3 protease (enzyme: 100 nM; inhibitor 50 lM; substrate 50 lM).
b
% Inhibition of the West Nile Virus (WNV) NS2B-NS3 protease (enzyme: 150 nM; inhibitor 50 lM; substrate 50 lM).
c
% Inhibition of thrombin (F2) (enzyme: 10 nM; inhibitor 25 lM; substrate 50 lM).
C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337 7323
Table 2
Activity of derivatives without acrylamide structure.
(COMT) inhibitor entacapone, we also tested this substance. Simi- 74 38.7 ± 1.1 46.5 ± 1.5 13.3 ± 3.2
2.3. Ki-values, target selectivities and aprotinin competition percentage inhibition against DEN and WNV protease (Table 1) and
assay the respective Ki-values (Table 3) can be explained by a lower Km-
value for the substrate of DEN protease. Compound 21 shows
The evaluation at the DEN and WNV NS2B-NS3 proteases was selectivity for the flaviviral proteases. In comparison to thrombin
performed using an established fluorescence assay with an inter- the selectivity ratio for this compound was 2.8:1 in favor of DEN
nally quenched FRET substrate as described by Steuer et al.13 For protease and 2.3:1 of WNV protease respectively. Quite interest-
the initial exploration of the new compounds and to search for lead ingly, compound 71 was identified as a thrombin inhibitor with a
structures, the evaluation was usually restricted to single-concen- Ki-value in the low micromolar range for thrombin and very low
tration experiments, performed in triplicate (percentage of inhibi- affinity for the flaviviral proteases.
tion at [inhibitor] = 50 lM). Selected compounds with pronounced The Ki-value of compound 21 at the DEN protease is in the range
activity or selectivity were assayed in more detail. Table 3 shows of various published peptidyl-aldehyde inhibitors for this target.3
the Ki-values and target selectivities of these compounds. Other non-peptidic inhibitors, like the recently published anthra-
The most active compound 21 has a Ki-value of 35.7 lM at DEN cene derivatives from Tomlinson et al., show activities in the
and 44.6 lM at WNV protease. The seeming contradiction between micromolar range.14 There might be a general difficulty to create
7324 C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337
Table 3
Ki-values and target selectivities of selected compounds.
a
Ki-value against DEN protease.
b
Ki-value against WNV protease.
c
Ki-value against thrombin (F2).
3. Conclusion
high affinities with small molecule inhibitors at the DEN protease 4. Experimental section
target.
The aprotinin competition assay described by Bodenreider et al. 4.1. Biological methods
can be used to identify compounds that bind to the active site of
DEN protease.15 The intrinsic fluorescence of Trp50, which is lo- 4.1.1. Substrate synthesis
cated near the transition of S1 to S2 pocket is quenched by UV- The internally quenched DEN NS2B-NS3 protease substrate
absorbing compounds binding to the active site. The fluorescence Abz-NleKRRS-3-(NO2)Y was synthesized by solid-phase synthesis
of Trp50 is partially restored if the compounds are displaced by on Rink amide resin according to the Fmoc-protocol. It was purified
the known competitive inhibitor aprotinin. Because of its advanta- by preparative HPLC using an ÄKTA Purifier, GE Germany, with a
geous spectroscopic properties (no inherent fluorescence at rele- RP-18 chromatography column (Lobar, Merck, Germany, 40–
vant wavelength and an absorption maximum near 330 nm), 63 lm, 25 310 mm). The mobile phase consisted of MeOH/0.1%
compound 8 with a Ki-value of 98.1 lM was used for this experi- TFA and H2O/0.1% TFA following a gradient of 20–100% MeOH in
ment. The results are shown in Figure 2 and indicate that aprotinin water with a flow rate of 1.5 ml/min. The internally quenched sub-
can at least partially displace compound 8 from the active site of strate for the WNV assay (Abz-GLKRGG-3-(NO2)Y) was synthesized
the enzyme. This indicates a direct interaction of the inhibitor 8 and purified as described above. The purity of both substrates was
with the catalytic center of the protein. assessed by HPLC and found to be higher than 95%. The identity
To exclude the possibility of an unspecific or promiscuous bind- was confirmed by MALDI-TOF-MS.
ing mode, the solutions (100 lM) of some of the most potent inhib-
itors in KH2PO4 buffer (pH 7) were analyzed by dynamic light 4.1.2. Expression and purification of DEN and WNV proteases
scattering (data not shown), without any hints of aggregate forma- The DEN NS2B-NS3 gene, with the hydrophilic cofactor NS2B
tion. In addition, the compounds were practically inactive against a connected to the protease domain NS3pro via a flexible glycine lin-
number of other, unrelated enzyme targets that are routinely ker, was synthesized by a commercial supplier. The expression
tested in our laboratory (Escherichia coli methionine aminopepti- plasmid pET28a (Novagen) with the inserted gene was used to
dase, Homo sapiens methionine aminopeptidase 1, E. coli MurA). transform E. coli BL 21 k (DE3) cells. Overnight cultures of the
C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337 7325
transformed cells were grown at 37 °C in standard LB-medium and 19F NMR spectra were recorded on Varian Mercury Plus
containing 50 lg/ml kanamycin. After the OD of 0.6–0.8 at (300 MHz) and Varian NMR System 500 (500 MHz) instruments
600 nm was reached, the expression was induced by addition of at 300 K in CDCl3, acetone-d6, DMSO-d6, CD3OD or D2O. Chemical
IPTG to a final concentration of 1 mM. The cells were grown at shifts given in parts per million (d, ppm) and the residuals of
30 °C for further 4 h and then collected by centrifugation at non-deuterated solvents were use as internal standard (1H NMR:
4500 g. The pellet was resuspended in buffer A (lysis buffer: CDCl3: d = 7.25 ppm, acetone-d6: d = 2.04 ppm, DMSO-d6:
50 mM Tris–HCl pH 7.9, 100 mM NaCl, 5% glycerol and 5 mM imid- d = 2.49 ppm, CD3OD: d = 3.30 ppm, D2O: d = 4.75 ppm; 13C NMR:
azole) and passed through a cell disruptor (One Shot, Constant Sys- CDCl3: d = 77.00 ppm, acetone-d6: d = 29.80 ppm, DMSO-d6:
tems). Afterwards the solution was centrifuged at 18,500g and 4 °C d = 39.50 ppm, CD3OD: d = 49.00 ppm). Coupling constants (J) are
for 40 min. The supernatant was then purified by Ni2+-affinity given in hertz (Hz). Multiplicity is reported as s (singlet), d (dou-
chromatography. The protein was eluted by increasing the imidaz- blet), t (triplet), quart (quartet), sept (septet), dd (doublet-doublet),
ole concentration from 5 mM to 250 mM. Stocks of purified protein ddd (doublet-doublet-doublet), dt (doublet-triplet), td (triplet-
were stored at 70 °C in 100 mM Tris–HCl pH 7.9, 50 mM NaCl and doublet), m (multiplet) and br (broad), respectively. IR spectra
50% glycerol. The expression and purification protocol for the WNV were recorded on a Jasco FT-IR spectrometer (FT/IR-4100) with a
protease was similar to the one given for DEN. Pike MIRacle ATR module and are reported in reciprocal centime-
ters (cm1). Mass spectra were measured on Finnigan MAT 8200
4.1.3. Flourimetric DEN and WNV protease assay (EI), Bruker micrOTOF-Q II (HR-ESI) and Bruker BIFLEX III (MAL-
The DEN protease assay was performed as described previously DI-TOF) instruments. Combustion elemental analysis was per-
(Steuer et al.).13 In short, continuous enzymatic assays were per- formed by double determination using a Foss Heraeus Vario EL
formed on a BMG Labtech Fluostar OPTIMA microtiter fluorescence analyzer. Flash chromatography was performed on a Biotage Isol-
plate reader using black 96 well V-bottom plates from Greiner. The era One purification system using silica gel (0.060–0.200 mm) car-
excitation wavelength was 320 nm and the emission was moni- tridges (KP-Sil) and UV monitoring. The reaction progress was
tored at 405 nm. The inhibitor concentration was 50 lM. The determined by thin layer chromatography on Merck Silica Gel
inhibitors were preincubated for 15 min with the enzyme plates 60 F254 (UV detection). Microwave synthesis was performed
(100 nM). Afterwards, the reaction was initiated by the addition using a Monowave 300 synthesis reactor from Anton Paar. Purity of
of the substrate to a final concentration of 50 lM. The activity of the compounds used in biological assays was determined by com-
the enzyme was determined as the slope per second (RFU/s) and bustion elemental analysis (to an accuracy of within ± 0.4%) and
monitored for 15 min. Experiments were performed in triplicate HPLC. HPLC was performed using an Agilent 1200 HPLC system
(n = 3) and the experimental values were averaged. The WNV pro- on a xTerra MS C18 (2.5 lm) 2.1 50 mm column. Detection was
tease assay was performed in analogy to the DEN protease assay. conducted at 285 and 320 nm. The system conditions were A:
Final concentration of the enzyme was 150 nM. H2O (0.1% TFA), B: CH3CN (0.1% TFA), flow rate: 0.3 ml/min, gradi-
ent: 10% B (1 min), 95% B (9 min), 95% B (11 min), 10% B (11.1 min),
4.1.4. Thrombin assay 10% B (20 min).
The thrombin assay was performed as a continuous fluorimetric
assay on a BMG Labtech Fluostar OPTIMA microtiter fluorescence 4.2.1. General procedure for the preparation of compounds 1, 5–
plate reader. The excitation wavelength was 355 nm and the emis- 15, 18–21, 23–33, 35–44, 46–54, 61, 62, 65, 67–69, 72–75, 92
sion wavelength was 460 nm. The protease was assayed against A solution of arylaldehyde (1 equiv), 2-cyanoacetamide or
the substrate Boc-VPR-AMC (Bachem, Germany). The final concen- derivative (1.0–1.5 equiv) and N-methylpiperazine (0.05–
trations of the enzyme and substrate were 10 and 50 lM respec- 1.05 equiv) in methanol (2–10 ml) was stirred at room tempera-
tively. The inhibitors were preincubated with the enzyme for ture overnight. After addition of an equivalent volume of water/
15 min at a concentration of 25 lM. The cleavage reaction was ini- methanol (1:1) or 1 N HCl/methanol (1:1) for acidic products, the
tiated by addition of the substrate. The assay buffer consisted of precipitate was collected by filtration and washed with water/
50 mM Tris–HCl pH 7.5, 150 mM NaCl and 0.05% Tween 20.17 methanol (1:1). If precipitation did not occur, the mixture was
The activity of the enzyme was determined as the slope per second evaporated and the residue was purified by flash chromatography.
(RFU/s) and monitored for 10 min.
4.2.1.1. (E)-2-Cyano-3-phenylacrylamide (1). Starting from benz-
4.1.5. Ki-value determination aldehyde (2.12 g, 20.0 mmol) and 2-cyanoacetamide (2.14 g,
Ki-values were determined by duplicate experiments using 25.5 mmol) compound 1 was obtained after precipitation as a col-
eight different substrate concentrations (25, 50, 75, 100, 150, orless solid (2.25 g, 65%). 1H NMR (300 MHz, acetone-d6): d = 7.20
200, 300, 400 lM) at three different inhibitor concentrations (25, (br m, 2H), 7.59 (m, 3H), 8.01 (m, 2H), 8.24 (s, 1H) ppm; 13C
50, 100 lM) and without inhibitor. Km-values of the different sub- NMR (75 MHz, acetone-d6): d = 106.5, 117.2, 130.0, 131.1, 133.0,
strates are 104 lM (DEN protease—Abz-NleKRRS-3-(NO2)Y), 133.2, 152.4, 162.9 ppm; IR (neat): 3395, 3154, 2219, 1686, 1595,
211 lM (WNV protease—Abz-GLKRGG-3-(NO2)Y) and 15.9 lM 1573, 1495, 1447, 1368, 1289, 1208, 1183, 1105, 952, 765, 741,
(thrombin—Boc-VPR-AMC). The calculation was done using Prism 682 cm1; MS (EI, 70 eV): m/z (%): 172.1 (77) [M]+; HRMS (ESI):
5.0 (GraphPad Software, Inc.). m/z [M+H]+ calcd for C10H9N2O: 173.0709, found: 173.0701. Anal.
Calcd for C10H8N2O: C, 69.76; H, 4.68; N, 16.27. Found: C, 69.64;
4.1.6. Tryptophan quenching assay H, 4.88; N, 16.26.
This assay was performed as described by Bodenreider et al.15
4.2.1.2. (E)-2-Cyano-3-(4-methylphenyl)acrylamide (5). Starting
4.2. Chemistry from 4-methylbenzaldehyde (240 mg, 2.0 mmol) and 2-cyanoacet-
amide (211 mg, 2.51 mmol) compound 5 was obtained after precip-
All chemicals were obtained from Sigma-Aldrich (Germany) and itation as a colorless solid (220 mg, 59%). 1H NMR (300 MHz,
Alfa Aesar, Johnson Matthey (Germany) and were of analytical acetone-d6): d = 2.41 (s, 3H), 7.13 (br m, 2H), 7.38 (m, 2H), 7.92 (m,
grade. No further purification steps were performed unless indi- 2H), 8.19 (s, 1H) ppm; 13C NMR (75 MHz, acetone-d6): d = 21.6,
cated. All solvents were used as obtained from the commercial 105.2, 117.5, 130.4, 130.7, 131.3, 144.3, 152.3, 162.9 ppm; IR (neat):
sources. Solvents were dried using standard procedures. 1H, 13C 3381, 3145, 2218, 1692, 1588, 1507, 1366, 1213, 1181, 1105,
7326 C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337
815 cm1; MS (EI, 70 eV): m/z (%): 186.1 (93) [M]+; HRMS (ESI): m/z (ESI): m/z [M+H]+ calcd for C19H20N3O: 306.1601, found:
[M+H]+ calcd for C11H11N2O: 187.0866, found: 187.0869. Anal. Calcd 306.1599; HPLC purity >96%.
for C11H10N2O: C, 70.95; H, 5.41; N, 15.04. Found: C, 70.86; H, 5.42; N,
15.00. 4.2.1.7. (E)-2-Cyano-3-[4-(hydroxymethyl)phenyl]acrylamide
(10). Starting from 93 (409 mg, 3.0 mmol) and 2-cyanoacetamide
4.2.1.3. (E)-2-Cyano-3-(4-isopropylphenyl)acrylamide (6). Start- (280 mg, 3.33 mmol) compound 10 was obtained after precipita-
ing from 4-isopropylbenzaldehyde (148 mg, 1.0 mmol) and 2-cya- tion as a colorless solid (420 mg, 69%). 1H NMR (300 MHz, ace-
noacetamide (93 mg, 1.11 mmol) compound 6 was obtained after tone-d6): d = 4.45 (t, J = 5.8 Hz, 1H), 4.73 (d, J = 5.8 Hz, 2H), 7.56
precipitation as a colorless solid (95 mg, 44%). 1H NMR (300 MHz, (m, 2H), 7.99 (m, 2H), 8.22 (s, 1H) ppm; 13C NMR (75 MHz, ace-
acetone-d6): d = 1.27 (d, J = 7.0 Hz, 6H), 3.01 (sept, J = 6.9 Hz, 1H), tone-d6): d = 64.1, 105.7, 117.4, 127.7, 131.2, 131.6, 148.6, 152.3,
7.10 (br m, 2H), 7.46 (m, 2H), 7.96 (m, 2H), 8.20 (s, 1H) ppm; 13C 162.8 ppm; IR (neat): 3501, 3328, 3153, 2218, 1697, 1592, 1382,
NMR (75 MHz, acetone-d6): d = 23.8, 34.9, 105.3, 117.5, 128.1, 1209, 1111, 1024, 1011, 956, 938, 829, 804, 679 cm1; HRMS
130.8, 131.5, 152.3, 155.0, 162.9 ppm; IR (neat): 3413, 3203, (ESI): m/z [M+H]+ calcd for C11H11N2O2: 203.0815, found:
2958, 2212, 1685, 1588, 1508, 1462, 1421, 1362, 1283, 1211, 203.0821. Anal. Calcd for C11H10N2O2: C, 65.34; H, 4.98; N, 13.85.
1189, 1053, 829 cm1; MS (EI, 70 eV): m/z (%): 214.1 (78) [M]+; Found: C, 65.47; H, 5.08; N, 13.68.
HRMS (ESI): m/z [M+H]+ calcd for C13H15N2O: 215.1179, found:
215.1183. Anal. Calcd for C13H14N2O: C, 72.87; H, 6.59; N, 13.07. 4.2.1.8. (E)-2-Cyano-3-(4-fluorophenyl)acrylamide (11). Starting
Found: C, 72.67; H, 6.66; N, 12.92. from 4-fluorobenzaldehyde (248 mg, 2.0 mmol) and 2-cyanoacet-
amide (215 mg, 2.56 mmol) compound 11 was obtained after pre-
4.2.1.4. (E)-2-Cyano-N-cyclopropyl-3-(4-isopropylphenyl)acryl- cipitation as a colorless solid (190 mg, 50%). 1H NMR (300 MHz,
amide (7). Starting from 4-isopropylbenzaldehyde (148 mg, acetone-d6): d = 7.15 (br m, 2H), 7.36 (m, 2H), 8.11 (m, 2H), 8.23
1.0 mmol) and 84 (137 mg, 1.10 mmol) compound 7 was obtained (s, 1H) ppm; 13C NMR (75 MHz, acetone-d6): d = 106.3,
after precipitation and recrystallization from methanol/acetone/ 117.0 + 117.3 (d, 2JCF = 22.2 Hz), 117.2, 129.7 + 129.7 (d,
water as a colorless solid (140 mg, 55%). 1H NMR (300 MHz, 4
JCF = 3.2 Hz), 133.8 + 133.9 (d, 3JCF = 9.2 Hz), 151.1, 162.6,
CDCl3): d = 0.64 (m, 2H), 0.88 (m, 2H), 1.26 (d, J = 6.9 Hz, 6H), 163.9 + 167.3 (d, 1JCF = 253 Hz) ppm; IR (neat): 3468, 3158, 2216,
2.87 (m, 1H), 2.96 (sept, J = 6.9 Hz, 1H), 6.42 (br m, 1H), 7.33 (m, 1697, 1585, 1502, 1414, 1381, 1301, 1234, 1163, 1113, 956,
2H), 7.86 (m, 2H), 8.29 (s, 1H) ppm; 13C NMR (75 MHz, CDCl3): 834 cm1; MS (EI, 70 eV): m/z (%): 190.1 (100) [M]+; HRMS (ESI):
d = 6.8, 23.5, 23.6, 34.3, 102.3, 117.3, 127.4, 129.5, 131.0, 152.9, m/z [M+H]+ calcd for C10H8FN2O: 191.0615, found: 191.0618. Anal.
154.7, 161.7 ppm; IR (neat): 3329, 3012, 2961, 2871, 2221, 1668, Calcd for C10H7FN2O: C, 63.16; H, 3.71; N, 14.73. Found: C, 62.98;
1592, 1558, 1511, 1420, 1363, 1273, 1204, 1096, 1014, 993, 959, H, 3.89; N, 14.57.
840, 824 cm1; MS (EI, 70 eV): m/z (%): 254.1 (40) [M]+; HRMS
(ESI): m/z [M+H]+ calcd for C16H19N2O: 255.1492, found: 4.2.1.9. (E)-2-Cyano-3-(2,4-dichlorophenyl)acrylamide (12).
255.1494. Anal. Calcd for C16H18N2O: C, 75.56; H, 7.13; N, 11.01. Starting from 2,4-dichlorobenzaldehyde (350 mg, 2.0 mmol) and
Found: C, 75.26; H, 7.20; N, 10.94. 2-cyanoacetamide (210 mg, 2.50 mmol) compound 12 was ob-
tained after precipitation as a colorless solid (252 mg, 52%). 1H
4.2.1.5. (E)-2-Cyano-3-(4-isopropylphenyl)-N-(tetrahydrofuran- NMR (300 MHz, acetone-d6): d = 7.31 (br m, 2H), 7.61 (ddd,
2-ylmethyl)acrylamide (8). Starting from 4-isopropylbenzalde- J = 8.5, 2.1, 0.6 Hz, 1H), 7.71 (d, J = 2.1 Hz, 1H), 8.13 (d,
hyde (148 mg, 1.0 mmol) and 85 (185 mg, 1.10 mmol) compound J = 8.5 Hz, 1H), 8.48 (t, J = 0.6 Hz, 1H) ppm; 13C NMR (75 MHz,
8 was obtained after precipitation and recrystallization from ace- acetone-d6): d = 111.0, 116.2, 128.9, 130.5, 130.7, 131.6, 136.9,
tone/water as a colorless solid (150 mg, 50%). 1H NMR (300 MHz, 138.6, 147.3, 161.8 ppm; IR (neat): 3388, 3168, 2229, 1708,
CDCl3): d = 1.26 (d, J = 6.9 Hz, 6H), 1.59 (m, 1H), 1.93 (m, 2H), 1604, 1584, 1469, 1379, 1206, 1144, 1108, 1050, 923, 862 cm1;
2.03 (m, 1H), 2.96 (sept, J = 6.9 Hz, 1H), 3.37 (m, 1H), 3.69 (m, MS (EI, 70 eV): m/z (%): 240.0 (6) [M]+; HRMS (ESI): m/z [M+H]+
1H), 3.79 (m, 1H), 3.91 (m, 1H), 4.05 (m, 1H), 6.68 (br s, 1H), calcd for C10H7Cl2N2O: 240.9930, found: 240.9927. Anal. Calcd
7.33 (m, 2H), 7.87 (m, 2H), 8.28 (s, 1H) ppm; 13C NMR (75 MHz, for C10H6Cl2N2O: C, 49.82; H, 2.51; N, 11.62. Found: C, 49.64; H,
CDCl3): d = 23.6, 25.9, 28.7, 34.3, 44.1, 68.3, 77.1, 102.7, 117.1, 2.75; N, 11.46.
127.4, 129.5, 130.9, 152.9, 154.6, 160.7 ppm; IR (neat): 3368,
2957, 2867, 2214, 1673, 1595, 1520, 1463, 1427, 1269, 1211, 4.2.1.10. (E)-3-(4-Bromophenyl)-2-cyanoacrylamide (13). Start-
1057, 918, 828 cm1; MS (EI, 70 eV): m/z (%): 298.1 (23) [M]+; ing from 4-bromobenzaldehyde (185 mg, 1.0 mmol) and 2-cyano-
HRMS (ESI): m/z [M+H]+ calcd for C18H23N2O2: 299.1754, found: acetamide (93 mg, 1.11 mmol) compound 13 was obtained after
299.1771. Anal. Calcd for C18H22N2O2: C, 72.46; H, 7.43; N, 9.39. precipitation as a colorless solid (235 mg, 94%). 1H NMR
Found: C, 72.05; H, 7.39; N, 9.35. (300 MHz, acetone-d6): d = 7.18 (br m, 2H), 7.78 (m, 2H), 7.94 (m,
2H), 8.20 (s, 1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 107.3,
4.2.1.6. (E)-2-Cyano-3-(4-isopropylphenyl)-N-(pyridin-2- 116.2, 125.9, 131.1, 131.7, 132.3, 149.3, 162.5 ppm; IR (neat):
ylmethyl)acrylamide (9). Starting from 4-isopropylbenzaldehyde 3437, 3142, 2215, 1694, 1600, 1578, 1488, 1374, 1205, 1185,
(148 mg, 1.0 mmol) and 86 (193 mg, 1.10 mmol) compound 9 1115, 1071, 1006, 954, 808 cm1; MS (EI, 70 eV): m/z (%): 249.9
was obtained after flash chromatography (cyclohexane/ethyl ace- (100) [M]+; HRMS (ESI): m/z [M+H]+ calcd for C10H8BrN2O:
tate) as a colorless solid (110 mg, 36%). 1H NMR (300 MHz, CDCl3): 250.9815, found: 250.9814. Anal. Calcd for C10H7BrN2O: C, 47.84;
d = 1.27 (d, J = 6.9 Hz, 6H), 2.97 (sept, J = 6.9 Hz, 1H), 4.72 (d, H, 2.81; N, 11.16. Found: C, 47.79; H, 2.98; N, 10.88.
J = 4.8 Hz, 2H), 7.22 (m, 1H), 7.28 (m, 1H), 7.34 (m, 2H), 7.68 (td,
J = 7.6, 1.8 Hz, 1H), 7.76 (br m, 1H), 7.89 (m, 2H), 8.32 (s, 1H), 4.2.1.11. (E)-3-(4-Bromophenyl)-2-cyano-N-cyclopropylacryla-
8.60 (m, 1H) ppm; 13C NMR (75 MHz, CDCl3): d = 23.6, 34.3, 45.1, mide (14). Starting from 4-bromobenzaldehyde (185 mg,
102.8, 117.1, 121.9, 122.6, 127.4, 129.6, 131.0, 136.8, 149.3, 1.0 mmol) and 84 (137 mg, 1.10 mmol) compound 14 was ob-
152.8, 154.6, 155.2, 160.6 ppm; IR (neat): 3370, 2956, 2217, tained after precipitation as a colorless solid with pure crystals
1663, 1590, 1523, 1472, 1434, 1413, 1288, 1270, 1209, 1194, used for X-ray analysis (50 mg, 17%). 1H NMR (300 MHz, acetone-
830, 749 cm1; MS (EI, 70 eV): m/z (%): 305.2 (9) [M]+; HRMS d6): d = 0.67 (m, 2H), 0.76 (m, 2H), 2.88 (m, 1H), 7.56 (br m, 1H),
C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337 7327
7.75 (m, 2H), 7.91 (m, 2H), 8.16 (s, 1H) ppm; 13C NMR (75 MHz, 4.2.1.16. (E)-2-Cyano-3-(4-hydroxyphenyl)acrylamide (21).
acetone-d6): d = 6.4, 24.3, 107.7, 116.7, 127.0, 132.3, 132.7, 133.2, Starting from 4-hydroxybenzaldehyde (611 mg, 5.0 mmol) and 2-
150.3, 162.3 ppm; IR (neat): 3325, 3016, 2221, 1669, 1582, 1509, cyanoacetamide (560 mg, 6.66 mmol) compound 21 was obtained
1274, 1197, 1074, 1009, 991, 835, 816 cm1; HRMS (ESI): m/z after precipitation as a pale yellow solid (850 mg, 90%). 1H NMR
[M+H]+ calcd for C13H12BrN2O: 291.0128, found: 291.0128. Anal. (300 MHz, acetone-d6): d = 7.00 (br m, 2H), 7.01 (m, 2H), 7.96 (m,
Calcd for C13H11BrN2O: C, 53.63; H, 3.81; N, 9.62. Found: C, 2H), 8.13 (s, 1H), 9.40 (br s, 1H) ppm; 13C NMR (75 MHz, DMSO-
53.62; H, 3.87; N, 9.59. d6): d = 101.5, 116.2, 117.3, 122.9, 132.9, 150.5, 161.7, 163.3 ppm;
IR (neat): 3449, 3362, 3159, 2227, 1651, 1599, 1569, 1511, 1412,
4.2.1.12. (E)-2-Cyano-3-(4-methoxyphenyl)acrylamide (15). 1374, 1287, 1229, 1179, 930, 835, 810 cm1; MS (EI, 70 eV): m/z
Starting from anisaldehyde (272 mg, 2.0 mmol) and 2-cyanoacet- (%): 188.1 (100) [M]+; HRMS (ESI): m/z [M+H]+ calcd for
amide (210 mg, 2.50 mmol) compound 15 was obtained after pre- C10H9N2O2: 189.0659, found: 189.0657. Anal. Calcd for
cipitation as a colorless solid (364 mg, 90%). 1H NMR (300 MHz, C10H8N2O2: C, 63.82; H, 4.28; N, 14.89. Found: C, 63.65; H, 4.40;
DMSO-d6): d = 3.84 (s, 3H), 7.12 (m, 2H), 7.72 (br m, 2H), 7.95 N, 14.68.
(m, 2H), 8.10 (s, 1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 55.6,
102.9, 114.8, 117.1, 124.4, 132.4, 150.1, 162.6, 163.1 ppm; IR 4.2.1.17. (E)-2-Cyano-N-cyclopropyl-3-(4-hydroxyphenyl)acryl-
(neat): 3444, 3166, 2208, 1693, 1579, 1508, 1385, 1364, 1309, amide (23). Starting from 4-hydroxybenzaldehyde (122 mg,
1259, 1176, 1024, 961, 824 cm1; MS (EI, 70 eV): m/z (%): 202.0 1.0 mmol) and 84 (135 mg, 1.09 mmol) compound 23 was ob-
(100) [M]+; HRMS (ESI): m/z [M+H]+ calcd for C11H11N2O2: tained after precipitation as a pale yellow solid (200 mg, 88%). 1H
203.0815, found: 203.0817. Anal. Calcd for C11H10N2O2: C, 65.34; NMR (300 MHz, acetone-d6): d = 0.66 (m, 2H), 0.73 (m, 2H), 2.87
H, 4.98; N, 13.85. Found: C, 65.21; H, 5.02; N, 13.73. (m, 1H), 7.00 (m, 2H), 7.36 (br m, 1H), 7.92 (m, 2H), 8.09 (s, 1H),
9.41 (br s, 1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 5.7, 23.4,
4.2.1.13. (E)-2-Cyano-3-(3-methoxyphenyl)acrylamide (18). 101.6, 116.2, 117.1, 122.9, 132.7, 149.9, 161.6, 163.0 ppm; IR
Starting from 3-methoxybenzaldehyde (272 mg, 2.0 mmol) and (neat): 3324, 3015, 2217, 1643, 1609, 1562, 1516, 1442, 1285,
2-cyanoacetamide (210 mg, 2.50 mmol) compound 18 was ob- 1201, 1161, 830 cm1; HRMS (ESI): m/z [M+H]+ calcd for
tained after precipitation as a colorless solid (198 mg, 49%). 1H C13H13N2O2: 229.0972, found: 229.0963. Anal. Calcd for
NMR (300 MHz, acetone-d6): d = 3.87 (m, 3H), 7.15 (m, 1H), C13H12N2O2: C, 68.41; H, 5.30; N, 12.27. Found: C, 68.34; H, 5.41;
7.19 (br m, 2H), 7.47 (m, 1H), 7.58 (m, 2H), 8.21 (s, 1H) ppm; N, 12.29.
13
C NMR (75 MHz, acetone-d6): d = 55.8, 106.8, 115.7, 117.3,
119.3, 123.7, 131.1, 134.3, 152.4, 160.9, 162.6 ppm; IR (neat): 4.2.1.18. (E)-2-Cyano-3-(4-hydroxyphenyl)-N-(tetrahydrofuran-
3453, 3331, 2213, 1611, 1574, 1497, 1468, 1447, 1426, 1371, 2-ylmethyl)acrylamide (24). Starting from 4-hydroxybenzalde-
1308, 1254, 1177, 1027, 956, 858, 778 cm1; MS (EI, 70 eV): hyde (183 mg, 1.50 mmol) and 85 (265 mg, 1.58 mmol) compound
m/z (%): 202.0 (100) [M]+; HRMS (ESI): m/z [M+H]+ calcd for 24 was obtained after precipitation as a colorless solid (360 mg,
C11H11N2O2: 203.0815, found: 203.0816. Anal. Calcd for 88%). 1H NMR (300 MHz, acetone-d6): d = 1.62 (m, 1H), 1.81–2.03
C11H10N2O2: C, 65.34; H, 4.98; N, 13.85. Found: C, 65.22; H, (m, 3H), 3.36 (m, 1H), 3.50 (m, 1H), 3.69 (m, 1H), 3.83 (m, 1H),
4.99; N, 13.79. 4.03 (m, 1H), 7.00 (m, 2H), 7.23 (br m, 1H), 7.95 (m, 2H), 8.14 (s,
1H), 9.45 (s, 1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 25.1,
4.2.1.14. (E)-2-Cyano-3-(3,4-dimethoxyphenyl)acrylamide (19). 28.5, 43.6, 67.1, 76.7, 101.1, 116.2, 117.2, 122.9, 132.8, 150.3,
Starting from 3,4-dimethoxybenzaldehyde (1.16 g, 6.98 mmol) and 161.6, 161.7 ppm; IR (neat): 3349, 3150, 2211, 1652, 1539, 1507,
2-cyanoacetamide (810 mg, 9.64 mmol) compound 19 was ob- 1440, 1286, 1210, 1169, 1069, 833 cm1; HRMS (ESI): m/z
tained after precipitation as a colorless solid (1.47 g, 90%). 1H [M+H]+ calcd for C15H17N2O3: 273.1234, found: 273.1234. Anal.
NMR (300 MHz, acetone-d6): d = 3.88 (s, 3H), 3.92 (s, 3H), 7.03 Calcd for C15H16N2O3: C, 66.16; H, 5.92; N, 10.29. Found: C,
(br m, 2H), 7.13 (d, J = 8.4 Hz, 1H), 7.60 (ddd, J = 8.4, 2.2, 0.5 Hz, 65.98; H, 5.90; N, 10.29.
1H), 7.75 (d, J = 2.2 Hz, 1H), 8.15 (s, 1H) ppm; 13C NMR (75 MHz,
acetone-d6): d = 56.1, 56.2, 102.7, 112.4, 113.2, 118.1, 125.7, 4.2.1.19. (E)-2-Cyano-3-(4-hydroxyphenyl)-N-(pyridin-2-yl-
126.9, 150.3, 152.4, 154.2, 163.1 ppm; IR (neat): 3386, 3186, methyl)acrylamide (25). Starting from 4-hydroxybenzaldehyde
3003, 2222. 1713, 1667, 1632, 1586, 1513, 1448, 1380, 1272, (183 mg, 1.50 mmol) and 86 (276 mg, 1.58 mmol) compound 25
1248, 1019, 956, 850 cm1; MS (EI, 70 eV): m/z (%): 232.2 (100) was obtained after precipitation as a colorless solid (260 mg,
[M]+; HRMS (ESI): m/z [M+H]+ calcd for C12H13N2O3: 233.0921, 62%). 1H NMR (300 MHz, DMSO-d6): d = 4.51 (d, J = 5.6 Hz, 2H),
found: 233.0923. Anal. Calcd for C12H12N2O3: C, 62.06; H, 5.21; 6.93 (m, 1H), 7.28 (m, 2H), 7.76 (td, J = 7.7, 1.7 Hz, 1H), 7.90 (m,
N, 12.06. Found: C, 62.01; H, 5.35; N, 12.00. 2H), 8.11 (s, 1H), 8.51 (m, 1H), 8.85 (m, 1H), 10.59 (s, 1H) ppm;
13
C NMR (75 MHz, DMSO-d6): d = 45.0, 100.8, 116.2, 117.2, 121.0,
4.2.1.15. (E)-2-Cyano-3-(3,4,5-trimethoxyphenyl)acrylamide 122.2, 122.9, 132.9, 136.7, 148.8, 150.7, 157.9, 161.7, 161.8 ppm;
(20). IR (neat): 3372, 2203, 1675, 1591, 1569, 1526, 1508, 1444, 1386,
Starting from 3,4,5-trimethoxybenzaldehyde (1.37 g, 6.98 mmol) 1297, 1244, 1202, 1171, 1011, 831 cm1; MS (EI, 70 eV): m/z (%):
and 2-cyanoacetamide (795 mg, 9.46 mmol) compound 20 was ob- 279.1 (17) [M]+; HRMS (ESI): m/z [M+H]+ calcd for C16H14N3O2:
tained after precipitation as a colorless solid (1.73 g, 94%). 1H NMR 280.1081, found: 280.1074. Anal. Calcd for C16H13N3O2: C, 68.81;
(300 MHz, acetone-d6): d = 3.83 (s, 3H), 3.89 (s, 6H), 7.14 (br m, H, 4.69; N, 15.05. Found: C, 68.49; H, 4.84; N, 15.02.
2H), 7.41 (s, 2H), 8.16 (s, 1H) ppm; 13C NMR (75 MHz, CDCl3):
d = 56.3, 61.1, 101.3, 108.3, 117.5, 126.7, 142.6, 153.3, 153.9, 4.2.1.20. (E)-3-(4-Hydroxyphenyl)-2-(morpholin-4-ylcarbon-
162.0 ppm; IR (neat): 3406, 3133, 2224, 1711, 1690, 1596, 1575, yl)acrylonitrile (26). Starting from 4-hydroxybenzaldehyde
1502, 1466, 1450, 1415, 1384, 1355, 1323, 1302, 1256, 1240, (183 mg, 1.50 mmol) and 87 (243 mg, 1.58 mmol) compound 26
1188, 1155, 1116, 991, 832 cm1; MS (EI, 70 eV): m/z (%): 262.2 was obtained after precipitation as a pale yellow solid (200 mg,
(100) [M]+; HRMS (ESI): m/z [M+H]+ calcd for C13H15N2O4: 52%). 1H NMR (300 MHz, acetone-d6): d = 3.67 (m, 8H), 6.99 (m,
263.1026, found: 263.1026. Anal. Calcd for C13H14N2O4: C, 59.54; 2H), 7.61 (s, 1H), 7.89 (m, 2H), 9.33 (s, 1H) ppm; 13C NMR
H, 5.38; N, 10.68. Found: C, 59.43; H, 5.50; N, 10.63. (75 MHz, acetone-d6): d = 67.1, 102.8, 116.9, 117.6, 125.2, 133.2,
7328 C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337
151.2, 162.1, 164.2 ppm; IR (neat): 3270, 2958, 2217 1634, 1588, tation and recrystallization from methanol/acetone/water (con-
1516, 1426, 1276, 1205, 1178, 1105, 1065, 1028, 1008, 839 cm1; taining hydrochloric acid) as a yellow solid (40 mg, 13%). 1H
HRMS (ESI): m/z [M+H]+ calcd for C14H15N2O3: 259.1077, found: NMR (300 MHz, acetone-d6): d = 1.21 (br m, 6H), 3.51 (br m,
259.1076. Anal. Calcd for C14H14N2O3: C, 65.11; H, 5.46; N, 10.85. 4H), 7.60 (s, 1H), 7.90 (d, J = 2.2 Hz, 1H), 8.18 (d, J = 2.2 Hz,
Found: C, 64.73; H, 5.59; N, 10.69. 1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 13.8, 24.7, 104.7,
116.2, 117.6, 118.4, 122.7, 137.2, 144.8, 147.1, 148.0,
4.2.1.21. (E)-3-(4-Hydroxyphenyl)-2-[(4-methylpiperazin-1-yl)- 162.6 ppm; IR (neat): 2983, 2210, 1629, 1607, 1537, 1440,
carbonyl]acrylonitrile hydrochloride (27). Starting from 4- 1310, 1244, 1139, 945, 871, 764 cm1; HRMS (ESI): m/z
hydroxybenzaldehyde (244 mg, 2.0 mmol) and 88 (360 mg, [M+H]+ calcd for C14H16N3O5: 306.1084, found: 306.1079; HPLC
2.15 mmol) the crude product was obtained after precipitation. purity >98%.
After resolving in methanol an excess of aqueous hydrochloric
acid was added, which resulted in precipitation of compound 4.2.1.26. (E)-2-Cyano-3-(4-hydroxy-3-methoxyphenyl)acrylam-
27 as a colorless solid (95 mg, 15%). 1H NMR (300 MHz, ide (32). Starting from vanillin (761 mg, 5.0 mmol) and 2-cyano-
CD3OD): d = 2.96 (s, 3H), 3.22 (br m, 2H), 3.45 (br m, 2H), acetamide (550 mg, 6.54 mmol) compound 32 was obtained after
3.57 (br m, 2H), 4.48 (br m, 2H), 6.90 (m, 2H), 7.75 (s, 1H), precipitation as a colorless solid (900 mg, 83%). 1H NMR
7.90 (m, 2H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 41.9, (300 MHz, acetone-d6): d = 3.91 (s, 3H), 6.99 (d, J = 8.4 Hz, 1H),
51.7, 99.6, 116.1, 117.0, 123.1, 132.5, 150.9, 161.7, 163.4 ppm; 7.02 (br m, 2H), 7.55 (ddd, J = 8.4, 2.2, 0.5 Hz, 1H), 7.76 (d,
IR (neat): 3069, 2671, 2596, 2464, 2201, 1659, 1606, 1576, J = 2.2 Hz, 1H), 8.13 (s, 1H), 8.73 (bs, 1H) ppm; 13C NMR (75 MHz,
1510, 1406, 1284, 1235, 1212, 1167, 976, 844 cm1; HRMS acetone-d6): d = 56.3, 101.9, 113.7, 116.5, 118.2, 125.1, 127.2,
(ESI): m/z [M+H]+ calcd for C15H18N3O2: 272.1394, found: 148.6, 152.2, 152.6, 163.3 ppm; IR (neat): 3469, 3364, 2211,
272.1394; HPLC purity >98%. 1681, 1563, 1508, 1433, 1404, 1285, 1163, 1130, 1018, 849 cm1;
MS (EI, 70 eV): m/z (%): 218.1 (100) [M]+; HRMS (ESI): m/z
4.2.1.22. (E)-2-Cyano-3-(3-hydroxyphenyl)acrylamide (28). [M+H]+ calcd for C11H11N2O3: 219.0764, found: 219.0760. Anal.
Starting from 3-hydroxybenzaldehyde (1.83 g, 15.0 mmol) and 2- Calcd for C11H10N2O3: C, 60.55; H, 4.62; N, 12.84. Found: C,
cyanoacetamide (1.61 g, 19.1 mmol) compound 28 was obtained 60.46; H, 4.72; N, 12.73.
after precipitation as a colorless solid (2.41 g, 85%). 1H NMR
(300 MHz, acetone-d6): d = 7.07 (m, 1H), 7.19, (br m 2H), 7.41 (m, 4.2.1.27. (E)-2-Cyano-3-(4-hydroxy-3-iodo-5-methoxyphenyl)-
2H), 7.52 (m, 1H), 8.15 (s, 1H), 8.89 (s, 1H) ppm; 13C NMR acrylamide (33). Starting from 4-hydroxy-3-iodo-5-methoxy-
(75 MHz, acetone-d6): d = 106.3, 115.9, 116.4, 119.7, 121.5, 130.3, benzaldehyde (1.39 g, 5.0 mmol) and 2-cyanoacetamide (526 mg,
133.1, 150.7, 157.8, 162.9 ppm; IR (neat): 3406, 3202, 2217, 6.26 mmol) compound 33 was obtained after precipitation and
1670, 1578, 1448, 1389, 1285, 1230, 1116, 996, 956, 862 cm1; recrystallization from acetone as a pale yellow solid (1.25 g, 73%).
MS (EI, 70 eV): m/z (%): 188.1 (100) [M]+; HRMS (ESI): m/z 1
H NMR (300 MHz, DMSO-d6): d = 3.85 (s, 3H), 7.66 (d, J = 2.0 Hz,
[M+H]+ calcd for C10H9N2O2: 189.0659, found: 189.0653; HPLC 1H), 7.76 (br m, 2H), 7.93 (d, J = 2.0 Hz, 1H), 8.03 (s, 1H), 10.60
purity >98%. (br m, 1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 56.1, 84.8,
103.0, 112.8, 117.1, 125.0, 133.9, 146.8, 149.3, 150.5, 162.9 ppm;
4.2.1.23. (E)-2-Cyano-3-(3,4-dihydroxyphenyl)acrylamide (29). IR (neat): 3459, 3343, 3323, 2215, 1669, 1572, 1551, 1494, 1473,
Starting from 3,4-dihydroxybenzaldehyde (415 mg, 3.0 mmol) 1415, 1328, 1302, 1263, 1246, 1163, 1130, 1035, 970, 844,
and 2-cyanoacetamide (333 mg, 3.96 mmol) compound 29 was ob- 761 cm1; MS (EI, 70 eV): m/z (%): 344.0 (100) [M]+; HRMS (ESI):
tained after precipitation as a pale brown solid (350 mg, 57%). 1H m/z [M+H]+ calcd for C11H10IN2O3: 344.9731, found: 344.9721;
NMR (300 MHz, acetone-d6): d = 6.98 (d, J = 8.4 Hz, 1H), 7.00 (br HPLC purity >97%.
m, 2H), 7.38 (ddd, J = 8.4, 2.2, 0.5 Hz, 1H), 7.68 (d, J = 2.2 Hz, 1H),
8.05 (s, 1H), 8.70 (bs, 1H), 8.90 (bs, 1H) ppm; 13C NMR (75 MHz, 4.2.1.28. (E)-2-Cyano-3-(4-nitrophenyl)acrylamide (35). Starting
acetone-d6): d = 101.6, 116.6, 116.8, 118.0, 125.2, 126.8, 146.3, from 4-nitrobenzaldehyde (151 mg, 51.0 mmol) and 2-cyanoacet-
151.1, 152.5, 163.4 ppm; IR (neat): 3461, 3410, 3313, 2603, 2208, amide (93 mg, 1.10 mmol) compound 35 was obtained after pre-
1675, 1567, 1446, 1391, 1293, 1262, 1194, 1167, 1122, 869, cipitation as a brown solid (120 mg, 55%). 1H NMR (300 MHz,
794 cm1; MS (EI, 70 eV): m/z (%): 204.1 (100) [M]+; HRMS (ESI): acetone-d6): d = 7.30 (br m, 2H), 8.23 (m, 2H), 8.36 (s, 1H), 8.42
m/z [M+H]+ calcd for C10H9N2O3: 205.0608, found: 205.0608. Anal. (m, 2H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 110.6, 115.7,
Calcd for C10H8N2O3: C, 58.82; H, 3.95; N, 13.72. Found: C, 58.70; H, 124.2, 131.0, 138.1, 148.3, 148.8, 162.1 ppm; IR (neat): 3429,
4.04; N, 13.58. 3340, 2224, 1686, 1591, 1378, 1341, 1295, 1201, 1106, 854,
768 cm1; MS (EI, 70 eV): m/z (%): 217.0 (100) [M]+; HRMS (ESI):
4.2.1.24. (E)-2-Cyano-3-(3,4-dihydroxy-5-nitrophenyl)acrylam- m/z [M+H]+ calcd for C10H8N3O3: 218.0560, found: 218.0558. Anal.
ide (30). Starting from 95 (183 mg, 1.0 mmol) and 2-cyanoacet- Calcd for C10H7N3O3: C, 55.30; H, 3.25; N, 19.35. Found: C, 55.14; H,
amide (110 mg, 1.31 mmol) compound 30 was obtained after 3.48; N, 19.03.
precipitation and recrystallization in acetone/water (containing
hydrochloric acid) as a yellow solid (150 mg, 60%). 1H NMR 4.2.1.29. (E)-2-Cyano-3-(4-cyanophenyl)acrylamide (36). Start-
(300 MHz, DMSO-d6): d = 7.60–8.00 (m, 4H), 8.04 (s, 1H), 10.88 ing from 4-formylbenzonitrile (131 mg, 1.0 mmol) and 2-cyano-
(br m, 2H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 104.7, 116.4, acetamide (105 mg, 1.25 mmol) compound 36 was obtained after
188.0, 119.2, 122.1, 137.3, 145.4, 148.1, 148.8, 162.8 ppm; IR precipitation as a colorless solid (95 mg, 48%). 1H NMR (300 MHz,
(neat): 3432, 3406, 3257, 2234, 1655, 1606, 1537, 1484, 1398, acetone-d6): d = 7.27 (br m, 2H), 7.98 (m, 2H), 8.16 (m, 2H), 8.30
1360, 1288, 1250, 1129, 941, 887, 866, 765 cm1; HRMS (ESI): m/ (s, 1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 110.0, 113.8,
z [M+H]+ calcd for C10H8N3O5: 250.0458, found: 250.0457; HPLC 115.8, 118.2, 130.3, 132.9, 136.3, 148.7, 162.1 ppm; IR (neat):
purity >98%. 3422, 3171, 2226, 1699, 1684, 1597, 1503, 1374, 1296, 1205,
1114, 1016, 936, 826 cm1; MS (EI, 70 eV): m/z (%): 197.0 (77)
4.2.1.25. (E)-2-Cyano-3-(3,4-dihydroxy-5-nitrophenyl)-N,N-di- [M]+; HRMS (ESI): m/z [M+H]+ calcd for C11H8N3O: 198.0662,
ethylacrylamide (31). Starting from 95 (183 mg, 1.0 mmol) and found: 198.0658. Anal. Calcd for C11H7N3O: C, 67.00; H, 3.58; N,
89 (175 mg, 1.25 mmol) compound 31 was obtained after precipi- 21.31. Found: C, 66.72; H, 3.79; N, 21.40.
C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337 7329
4.2.1.30. 4-[(E)-3-Amino-2-cyano-3-oxoprop-1-en-1-yl]benzoic for C15H18N3O: 256.1444, found: 216.1458. Anal. Calcd for
acid (37). Starting from 4-formylbenzoic acid (2.40 g, 16.0 mmol) C15H17N3O: C, 70.56; H, 6.71; N, 16.46. Found: C, 70.25; H, 6.78;
and 2-cyanoacetamide (1.48 g, 17.6 mmol) compound 37 was ob- N, 16.31.
tained after precipitation as a colorless solid (2.98 g, 86%). 1H
NMR (300 MHz, DMSO-d6): d = 7.83 (bs, 1H), 7.99 (m, 3H), 8.07 4.2.1.35. (E)-2-Cyano-3-[4-(dimethylamino)phenyl]-N-(tetrahy-
(m, 2H), 8.23 (s, 1H) ppm; 13C NMR (75 MHz, DMSO-d6): drofuran-2-ylmethyl)acrylamide (42). Starting from 4-(dimeth-
d = 108.9, 116.0, 129.9, 130.0, 133.4, 135.8, 149.4, 162.4, ylamino)benzaldehyde (373 mg, 2.50 mmol) and 85 (459 mg,
166.5 ppm; IR (neat): 3389, 3218, 2227, 1709, 1677, 1620, 1581, 2.73 mmol) compound 42 was obtained after precipitation as an
1416, 1281, 1208, 1193, 1119, 1103, 974, 856, 769 cm1; MS (EI, orange solid (681 mg, 91%). 1H NMR (300 MHz, acetone-d6):
70 eV): m/z (%): 216.1 (100) [M]+; HRMS (ESI): m/z [M+H]+ calcd d = 1.62 (m, 1H), 1.82–2.00 (m, 3H), 3.11 (s, 6H), 3.36 (m, 1H),
for C11H9N2O3: 217.0608, found: 217.0611; HPLC purity >98%. 3.49 (m, 1H), 3.68 (m, 1H), 3.83 (m, 1H), 4.02 (m, 1H), 6.83 (m,
2H), 7.04 (br m, 1H), 7.91 (m, 2H), 8.05 (s, 1H) ppm; 13C NMR
4.2.1.31. 4-[(E)-2-Cyano-3-(cyclopropylamino)-3-oxoprop-1-en- (75 MHz, acetone-d6): d = 26.3, 40.0, 44.6, 68.4, 78.1, 97.4, 110.9,
1-yl]benzoic acid (38). Starting from 4-formylbenzoic acid 112.4, 119.0, 120.3, 133.8, 152.0, 154.3, 162.4 ppm; IR (neat):
(380 mg, 2.53 mmol) and 84 (335 mg, 2.69 mmol) compound 38 3409, 2973, 2927, 2870, 2832, 2194, 1651, 1610, 1569, 1505,
was obtained after precipitation and recrystallization from ace- 1444, 1378, 1324, 1285, 1234, 1176, 1080, 1010, 945, 817 cm1;
tone/water (containing hydrochloric acid) as a colorless solid MS (EI, 70 eV): m/z (%): 299.1 (86) [M]+; HRMS (ESI): m/z [M+H]+
(260 mg, 40%). 1H NMR (300 MHz, DMSO-d6): d = 0.59 (m, 2H), calcd for C17H22N3O2: 300.1707, found: 300.1719. Anal. Calcd for
0.69 (m, 2H), 2.78 (m, 1H), 7.98 (m, 2H), 8.07 (m, 2H), 8.14 (s, C17H21N3O2: C, 68.20; H, 7.07; N, 14.04. Found: C, 68.04; H, 7.15;
1H), 8.58 (d, J = 4.0 Hz, 1H), 13.27 (s, 1H) ppm; 13C NMR N, 13.91.
(75 MHz, DMSO-d6): d = 5.7, 12.4, 108.8, 115.9, 129.9, 129.9
133.4, 135.8, 148.9, 162.1, 166.5 ppm; IR (neat): 2833, 2217, 4.2.1.36. (E)-2-Cyano-3-[4-(dimethylamino)phenyl]-N-(pyridin-
1681, 1607, 1575, 1505, 1427, 1288, 1249, 1203, 1108, 1013, 2-ylmethyl)acrylamide (43). Starting from 4-(dimethyl-
928, 851 cm1; HRMS (ESI): m/z [M+H]+ calcd for C14H13N2O3: amino)benzaldehyde (150 mg, 1.0 mmol) and 86 (193 mg,
257.0921, found: 257.0916; HPLC purity >97%. 1.10 mmol) compound 43 was obtained after precipitation as
an orange solid (300 mg, 98%). 1H NMR (300 MHz, DMSO-d6):
4.2.1.32. 3-[(E)-2-cyano-3-(cyclopropylamino)-3-oxoprop-1-en- d = 3.05 (s, 6H), 4.50 (d, J = 5.7 Hz, 2H), 6.82 (m, 2H), 7.25 (m,
1-yl]benzoic acid (39). Starting from 3-formylbenzoic acid 1H), 7.31 (d, J = 7.7 Hz, 1H), 7.75 (td, J = 7.7, 1.8 Hz, 1H), 7.88
(250 mg, 1.67 mmol) and 2-cyanoacetamide (227 mg, 2.70 mmol) (m, 2H), 8.03 (s, 1H), 8.50 (m, 1H), 8.66 (m, 1H) ppm; 13C
compound 39 was obtained after precipitation as a colorless solid NMR (75 MHz, DMSO-d6): d = 39.5, 44.9, 96.4, 111.6, 118.2,
(194 mg, 45%). 1H NMR (300 MHz, acetone-d6): d = 0.68 (m, 2H), 118.7, 120.9, 122.1, 132.8, 136.7, 148.8, 150.7, 153.0, 158.1,
0.76 (m, 2H), 2.89 (m, 1H), 7.61 (br m, 1H), 7.71 (t, J = 7.8 Hz, 162.3 ppm; IR (neat): 3377, 2913, 2196, 1649, 1609, 1565,
1H), 8.20 (m, 2H), 8.27 (s, 1H), 8.62 (m, 1H) ppm; 13C NMR 1503, 1440, 1415, 1374, 1289, 1169, 995, 813, 749 cm1; MS
(75 MHz, acetone-d6): d = 6.4, 24.2, 108.4, 116.5, 130.4, 132.0, (EI, 70 eV): m/z (%): 306.1 (71) [M]+; HRMS (ESI): m/z [M+Na]+
132.4, 133.6, 133.6, 134.9, 150.6, 166.6 ppm; IR (neat): 3350, calcd for C18H18N4NaO: 329.1373, found: 329.1365. Anal. Calcd
2222, 1675, 1597, 1509, 1457, 1415, 1362, 1286, 1203, 1170, for C18H18N4O: C, 70.57; H, 5.92; N, 18.29. Found: C, 70.26; H,
933, 750 cm1; MS (EI, 70 eV): m/z (%): 256.0 (80) [M]+; HRMS 6.03; N, 18.33.
(ESI): m/z [M+H]+ calcd for C14H13N2O3: 257.0921, found:
257.0919; HPLC purity >98%. 4.2.1.37. (E)-3-[4-(Dimethylamino)phenyl]-2-(morpholin-4-yl-
carbonyl)acrylonitrile (44). Starting from 4-(dimethyl-
4.2.1.33. (E)-2-Cyano-3-[4-(dimethylamino)phenyl]acrylamide amino)benzaldehyde (300 mg, 2.01 mmol) and 87 (310 mg,
(40). Starting from 4-(dimethylamino)benzaldehyde (1.52 g, 2.01 mmol) compound 44 was obtained after precipitation as a yel-
10.2 mmol) and 2-cyanoacetamide (850 mg, 10.2 mmol) com- low solid (250 mg, 44%). 1H NMR (300 MHz, acetone-d6): d = 3.09
pound 40 was obtained after precipitation as an orange solid (s, 6H), 3.65 (m, 8H), 6.80 (m, 2H), 7.57 (s, 1H), 7.87 (m, 2H)
(1.62 g, 75%). 1H NMR (300 MHz, acetone-d6): d = 3.11 (s, 6H), ppm; 13C NMR (75 MHz, acetone-d6): d = 40.0, 46.5, 67.1, 98.3,
6.83 (m, 2H), 6.84 (br m, 2H), 7.92 (m, 2H), 8.04 (s, 1H) ppm; 13C 112.3, 118.6, 120.7, 133.2, 152.2, 154.0, 165.1 ppm; IR (neat):
NMR (75 MHz, acetone-d6): d = 40.0, 97.3, 112.4, 119.1, 120.2, 2954, 2911, 2862, 2197, 1633, 1604, 1561, 1521, 1425, 1363,
133.8, 152.4, 154.3, 164.1 ppm; IR (neat): 3398, 3143, 2198, 1323, 1266, 1232, 1169, 1111, 1021, 950, 827 cm1; MS (EI,
1676, 1608, 1558, 1518, 1440, 1358, 1235, 1166, 943, 808 cm1; 70 eV): m/z (%): 285.1 (100) [M]+; HRMS (ESI): m/z [M+H]+ calcd
MS (EI, 70 eV): m/z (%): 215.1 (100) [M]+; HRMS (ESI): m/z for C16H20N3O2: 286.1550, found: 286.1557. Anal. Calcd for
[M+H]+ calcd for C12H14N3O: 216.1131, found: 216.1137. Anal. C16H19N3O2: C, 67.35; H, 6.71; N, 14.73. Found: C, 67.26; H, 6.71;
Calcd for C12H13N3O: C, 66.96; H, 6.09; N, 19.52. Found: C, 66.61; N, 14.59.
H, 6.35; N, 19.18.
4.2.1.38. (E)-2-Cyano-3-pyridin-4-ylacrylamide (46). Starting
4.2.1.34. (E)-2-Cyano-N-cyclopropyl-3-[4-(dimethylamino)- from isonicotinaldehyde (421 mg, 3.93 mmol) and 2-cyanoacet-
phenyl]acrylamide (41). Starting from 4-(dimethylamino)benzal- amide (430 mg, 5.12 mmol) compound 46 was obtained after pre-
dehyde (450 mg, 3.02 mmol) and 84 (395 mg, 3.18 mmol) cipitation as a colorless solid (235 mg, 35%). 1H NMR (300 MHz,
compound 41 was obtained after precipitation as a yellow solid acetone-d6): d = 7.30 (br m, 2H), 7.82 (m, 2H), 8.22 (s, 1H), 8.80
(590 mg, 77%). 1H NMR (300 MHz, acetone-d6): d = 0.64 (m, 2H), (m, 2H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 111.5, 115.5,
0.72 (m, 2H), 2.85 (m, 1H), 3.11 (s, 6H), 6.82 (m, 2H), 7.16 (br m, 122.9, 139.1, 148.2, 150.7, 161.9 ppm; IR (neat): 3426, 3027,
1H), 7.89 (m, 2H), 8.01 (s, 1H) ppm; 13C NMR (75 MHz, DMSO- 2216, 1696, 1639, 1598, 1546, 1415, 1359, 1239, 1204, 1059,
d6): d = 5.7, 23.3, 39.5, 97.4, 111.6, 118.0, 118.7, 132.5, 149.8, 1004, 974, 853, 806 cm1; MS (EI, 70 eV): m/z (%): 173.1 (100)
152.8, 163.6 ppm; IR (neat): 3294, 2915, 2207, 1606, 1571, 1525, [M]+; HRMS (ESI): m/z [M+H]+ calcd for C9H8N3O: 174.0662, found:
1437, 1382, 1361, 1328, 1286, 1254, 1174, 950, 825 cm1; MS 174.0653. Anal. Calcd for C9H7N3O: C, 62.42; H, 4.07; N, 24.27.
(EI, 70 eV): m/z (%): 255.1 (54) [M]+; HRMS (ESI): m/z [M+H]+ calcd Found: C, 62.24; H, 4.17; N, 24.21.
7330 C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337
4.2.1.39. (E)-2-Cyano-3-pyridin-3-ylacrylamide (47). Starting 137.4, 138.5, 142.4, 161.9 ppm; IR (neat): 3317, 3064, 3009,
from nicotinaldehyde (214 mg, 2.0 mmol) and 2-cyanoacetamide 2219, 1659, 1578, 1517, 1502, 1415, 1265, 1242, 1205, 1165,
(210 mg, 2.50 mmol) compound 47 was obtained after precipitation 1098, 1046, 1018, 977, 955, 852, 809 cm1; HRMS (ESI): m/z
as a colorless solid (205 mg, 59%). 1H NMR (300 MHz, DMSO-d6): [M+H]+ calcd for C11H10BrN2OS: 296.9692, found: 296.9696. Anal.
d = 7.60 (dd, J = 8.1, 4.8 Hz, 1H), 7.85 (br m, 1H), 7.98 (br m, 1H), Calcd for C11H9BrN2OS: C, 44.46; H, 3.05; N, 9.43. Found: C,
8.23 (s, 1H), 8.36 (m, 1H), 8.72 (dd, J = 4.8, 1.6 Hz, 1H), 8.97 (m, 44.39; H, 3.29; N, 9.47.
1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 108.9, 116.1, 124.1,
128.1, 135.9, 147.7, 151.2, 152.3, 162.1 ppm; IR (neat): 3127, 4.2.1.44. (E)-3-(3-Bromo-2-thienyl)-2-cyanoacrylamide (52).
2217, 1712, 1675, 1637, 1593, 1490, 1414, 1371, 1255, 1223, 1193, Starting from 3-bromothiophene-2-carbaldehyde (192 mg,
1137, 1097, 1028, 967, 803 cm1; MS (EI, 70 eV): m/z (%): 173.1 1.0 mmol) and 2-cyanoacetamide (105 mg, 1.25 mmol) compound
(85) [M]+; HRMS (ESI): m/z [M+H]+ calcd for C9H8N3O: 174.0662, 52 was obtained after precipitation as a pale brown solid (220 mg,
found: 174.0662. Anal. Calcd for C9H7N3O: C, 62.42; H, 4.07; N, 86%). 1H NMR (300 MHz, acetone-d6): d = 7.19 (br m, 2H), 7.38 (d,
24.27. Found: C, 62.18; H, 4.18; N, 24.11. J = 5.4 Hz, 1H), 8.12 (dd, J = 5.4, 1.0 Hz, 1H), 8.47 (d, J = 1.0 Hz,
1H) ppm; 13C NMR (75 MHz, acetone-d6): d = 104.3, 116.9, 122.8,
4.2.1.40. (E)-2-Cyano-3-(2-thienyl)acrylamide (48). Starting from 131.7, 132.3, 134.9, 142.4, 162.2 ppm; IR (neat): 3468, 3114,
thiophene-2-carbaldehyde (337 mg, 3.0 mmol) and 2-cyanoacet- 2211, 1696, 1615, 1581, 1475, 1412, 1374, 1304, 1236, 1115,
amide (333 mg, 3.96 mmol) compound 48 was obtained after pre- 937, 876, 735 cm1; MS (EI, 70 eV): m/z (%): 256.0 (2) [M]+; HRMS
cipitation as a pale orange solid (365 mg, 58%). 1H NMR (300 MHz, (ESI): m/z [M+H]+ calcd for C8H6BrN2OS: 256.9379, found:
acetone-d6): d = 7.11 (br m, 2H), 7.31 (m, 1H), 7.90 (m, 1H), 8.01 256.9376. Anal. Calcd for C8H5BrN2OS: C, 37.37; H, 1.96; N,
(m, 1H), 8.40 (m, 1H) ppm; 13C NMR (75 MHz, acetone-d6): 10.90. Found: C, 37.46; H, 2.11; N, 10.82.
d = 102.4, 117.4, 129.3, 135.1, 137.2, 138.1, 144.9, 162.8 ppm; IR
(neat): 3467, 3296, 3133, 2206, 1696, 1616, 1573, 1417, 1375, 4.2.1.45. (E)-3-(5-Bromo-2-furyl)-2-cyano-N-cyclopropylacryla-
1355, 1301, 1242, 1215, 1107, 1081, 1048, 858, 738 cm1; MS (EI, mide (53). Starting from 5-bromo-2-furaldehyde (528 mg,
70 eV): m/z (%): 178.1 (100) [M]+; HRMS (ESI): m/z [M+H]+ calcd 3.0 mmol) and 84 (410 mg, 3.30 mmol) compound 53 was ob-
for C8H7N2OS: 179.0274, found: 179.0273. Anal. Calcd for tained after precipitation as a yellow solid (410 mg, 49%). 1H
C8H6N2OS: C, 53.92; H, 3.39; N, 15.72. Found: C, 53.77; H, 3.52; N, NMR (300 MHz, acetone-d6): d = 0.67 (m, 2H), 0.74 (m, 2H), 2.86
15.59. (m, 1H), 6.83 (dd, J = 3.7, 0.3 Hz, 1H), 7.35 (dd, J = 3.7, 0.6 Hz, 1H),
7.45 (br m, 1H), 7.90 (m, 1H) ppm; 13C NMR (75 MHz, DMSO-d6):
4.2.1.41. (E)-3-(5-Chloro-2-thienyl)-2-cyanoacrylamide (49). d = 5.7, 23.4, 101.5, 115.7, 116.0, 123.3, 128.8, 134.0, 150.3,
Starting from 5-chlorothiophene-2-carbaldehyde (160 mg, 162.0 ppm; IR (neat): 3306, 3105, 3039, 2225, 1662, 1605, 1517,
1.10 mmol) and 2-cyanoacetamide (115 mg, 1.37 mmol) com- 1453, 1362, 1283, 1262, 1200, 1091, 1019, 946, 927, 803 cm1;
pound 49 was obtained after precipitation as a pale brown solid HRMS (ESI): m/z [M+H]+ calcd for C11H10BrN2O2: 280.9920, found:
(110 mg, 47%). 1H NMR (300 MHz, acetone-d6): d = 7.11 (br m, 280.9911. Anal. Calcd for C11H9BrN2O2: C, 47.00; H, 3.23; N, 9.97.
2H), 7.28 (d, J = 4.1 Hz, 1H), 7.76 (d, J = 4.1 Hz, 1H), 8.30 (s, 1H) Found: C, 46.79; H, 3.43; N, 9.96.
ppm; 13C NMR (75 MHz, DMSO-d6): d = 102.4, 116.5, 128.4,
134.8, 136.7, 138.1, 143.2, 162.1 ppm; IR (neat): 3393, 3159, 4.2.1.46. (E)-2-Cyano-3-(1H-indol-3-yl)acrylamide (54). Starting
2207, 1683, 1583, 1509, 1422, 1377, 1290, 1246, 1210, 1071, from 1H-indole-3-carbaldehyde (726 mg, 5.0 mmol) and 2-cyano-
1005, 943, 804 cm1; MS (EI, 70 eV): m/z (%): 212.1 (57) [M]+; acetamide (535 mg, 6.37 mmol) compound 54 was obtained after
HRMS (ESI): m/z [M+H]+ calcd for C8H6ClN2OS: 212.9884, found: precipitation as a pale yellow solid (700 mg, 66%). 1H NMR
212.9879. Anal. Calcd for C8H5ClN2OS: C, 45.18; H, 2.37; N, 13.17. (300 MHz, acetone-d6): d = 6.90 (br m, 2H), 7.28 (m, 2H), 7.59 (m,
Found: C, 45.12; H, 2.53; N, 13.04. 1H), 7.95 (m, 1H), 8.58 (m, 2H) ppm; 13C NMR (75 MHz, acetone-
d6): d = 98.1, 111.3, 113.4, 119.1, 119.4, 122.6, 124.3, 128.5,
4.2.1.42. (E)-3-(5-Bromo-2-thienyl)-2-cyanoacrylamide (50). 131.0, 137.3, 144.0, 164.1 ppm; IR (neat): 3471, 3371, 3110,
Starting from 5-bromothiophene-2-carbaldehyde (382 mg, 2210, 1684, 1584, 1563, 1512, 1485, 1459, 1422, 1373, 1329,
2.0 mmol) and 2-cyanoacetamide (215 mg, 2.56 mmol) compound 1298, 1228, 1130, 1112, 1061, 752 cm1; MS (EI, 70 eV): m/z (%):
50 was obtained after precipitation as a pale yellow solid (150 mg, 211.1 (100) [M]+; HRMS (ESI): m/z [M+H]+ calcd for C12H10N3O:
29%). 1H NMR (300 MHz, acetone-d6): d = 7.12 (br m, 2H), 7.40 (d, 212.0818, found: 212.0819. Anal. Calcd for C12H9N3O: C, 68.24;
J = 4.1 Hz, 1H), 7.71 (dd, J = 4.1, 0.6 Hz, 1H), 8.32 (d, J = 0.6 Hz, H, 4.29; N, 19.89. Found: C, 68.00; H, 4.55; N, 19.67.
1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 102.6, 116.6, 121.2,
131.9, 137.4, 138.6, 142.9, 162.2 ppm; IR (neat): 3426, 3170, 4.2.1.47. (E)-2-Cyano-3-[2,3,5,6-tetrafluoro-4-(4-methylpipera-
2209, 1682, 1578, 1504, 1413, 1365, 1283, 1238, 1204, 1113, zin-1-yl)phenyl]acrylamide (61). Starting from Pentafluorobenz-
1077, 1058, 976, 941, 805 cm1; HRMS (ESI): m/z [M+H]+ calcd aldehyde (197 mg, 1.0 mmol), 2-cyanoacetamide (105 mg,
for C8H6BrN2OS: 256.9379, found: 256.9378. Anal. Calcd for 1.25 mmol) and N-methylpiperazine (0.11 ml, 1.0 mmol) com-
C8H5BrN2OS: C, 37.37; H, 1.96; N, 10.90. Found: C, 37.05; H, 2.22; pound 61 was obtained after precipitation and recrystallization
N, 10.80. from methanol/water as a pale yellow solid (12 mg, 4%). 1H NMR
(300 MHz, CD3OD): d = 2.35 (s, 3H), 2.59 (m, 4H), 3.44 (m, 4H),
4.2.1.43. (E)-3-(5-Bromo-2-thienyl)-2-cyano-N-cyclopropyl- 8.07 (m, 1H) ppm; 19F NMR (282 MHz, CD3OD): d = 153.2,
acrylamide (51). Starting from 5-bromothiophene-2-carbalde- 138.6 ppm; 13C NMR (125 MHz, DMSO-d6): d = 45.8, 50.1, 54.8,
hyde (1.0 g, 5.23 mmol) and 84 (720 mg, 5.80 mmol) compound 103.4, 114.1, 114.8, 132.7, 136.9 139.9, 141.9, 143.6, 145.6,
51 was obtained after precipitation as a pale yellow solid (1.48 g, 161.3 ppm; IR (neat): 3500, 3400, 2806, 2226, 1702, 1643, 1604,
95%). 1H NMR (300 MHz, DMSO-d6): d = 0.58 (m, 2H), 0.66 (m, 1486, 1455, 1384, 1351, 1289, 1273, 1187, 1160, 1140, 980 cm1;
2H), 2.76 (m, 1H), 7.45 (d, J = 4.1 Hz, 1H), 7.68 (dd, J = 4.1, 0.4 Hz, MS (EI, 70 eV): m/z (%): 342.4 (51) [M]+; HRMS (ESI): m/z [M+H]+
1H), 8.26 (d, J = 0.4 Hz, 1H), 8.41 (br m, 2H) ppm; 13C NMR calcd for C15H15F4N4O: 343.1177, found: 343.1175; HPLC purity
(75 MHz, DMSO-d6): d = 5.7, 23.4, 102.5, 116.4, 121.0, 131.8, >98%.
C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337 7331
4.2.1.48. (E)-2-Cyano-N-cyclopropyl-3-(3-morpholin-4-ylphe- 2H), 6.99 (dd, J = 15.0, 11.7 Hz, 1H), 7.22 (br m, 1H), 7.32 (d,
nyl)acrylamide (62). Starting from 97 (75 mg, 0.39 mmol) and J = 15.0 Hz, 1H), 7.55 (m, 2H), 7.91 (d, J = 11.7 Hz, 1H) ppm; 13C
84 (62 mg, 0.5 mmol) compound 62 was obtained after flash chro- NMR (75 MHz, acetone-d6): d = 6.5, 24.1, 40.1, 103.4, 112.8,
matography (cyclohexane/ethyl acetate) as a yellow oil (18 mg, 116.6, 118.5, 118.6, 123.7, 131.0, 149.2, 153.2, 153.5 ppm; IR
16%). 1H NMR (300 MHz, CDCl3): d = 0.65 (m, 2H), 0.89 (m, 2H), (neat): 3325, 2206, 1663, 1588, 1504, 1443, 1369, 1284, 1231,
2.86 (m, 1), 3.20 (m, 4H), 3.87 (m, 4H), 6.42 (br m, 1H), 7.07 (m, 1193, 1151, 987, 942, 811 cm1; MS (EI, 70 eV): m/z (%): 281.1
1H), 7.36 (m, 2H), 7.47 (m, 1H), 8.29 (s, 1H) ppm; 13C NMR (56) [M]+; HRMS (ESI): m/z [M+H]+ calcd for C17H20N3O:
(75 MHz, CDCl3): d = 6.9, 23.6, 48.8, 66.7, 103.4, 116.4, 117.2, 282.1601, found: 282.1605. Anal. Calcd for C17H19N3O: C, 72.57;
120.0, 122.6, 129.9, 132.6, 151.6, 153.7, 161.5 ppm; IR (neat): H, 6.81; N, 14.94. Found: C, 72.45; H, 6.78; N, 14.74.
2962, 2850, 2210, 1671, 1600, 1522, 1447, 1263, 1242, 1118,
1069, 1027, 996, 887, 780, 683 cm1; HRMS (ESI): m/z [M+H]+ calcd 4.2.1.53. (E)-3-[4-(Benzyloxy)-3-methoxyphenyl]-2-cyanoacryl-
for C17H20N3O2: 298.1550, found: 298.1548; HPLC purity >93%. amide (72). Starting from 98 (242 mg, 1.0 mmol) and 2-cyano-
acetamide (105 mg, 1.25 mmol) compound 72 was obtained after
4.2.1.49. (E)-2-Cyano-N-cyclopropyl-3-[4-(morpholin-4-ylcar- precipitation as a colorless solid (190 mg, 62%). 1H NMR
bonyl)phenyl]acrylamide (65). Starting from 96 (220 mg, (300 MHz, acetone-d6): d = 3.90 (s, 3H), 5.26 (s, 2H), 7.22 (d,
1.0 mmol) and 84 (137 mg, 1.10 mmol) compound 65 was ob- J = 8.4 Hz, 1H), 7.31–7.45 (m, 3H), 7.50 (m, 1H), 7.52 (m, 1H),
tained after flash chromatography (cyclohexane/ethyl acetate) as 7.59 (ddd, J = 8.4, 2.2, 0.5 Hz, 1H), 7.78 (d, J = 2.2 Hz, 1H), 8.15 (s,
a pale yellow oil (70 mg, 22%). 1H NMR (300 MHz, CDCl3): 1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 55.5, 69.9, 103.0,
d = 0.66 (m, 2H), 0.90 (m, 2H), 2.89 (m, 1H), 3.34–3.86 (br m, 112.6, 113.2, 117.2, 124.7, 125.2, 128.0, 128.1, 128.5, 136.3,
8H), 6.44 (br m, 1H), 7.51 (m, 2H), 7.95 (m, 2H), 8.33 (s, 1H) 148.9, 150.5, 151.5, 163.0 ppm; IR (neat): 3166, 2209, 1659,
ppm; 13C NMR (75 MHz, CDCl3): d = 6.9, 23.7, 38.6, 66.8, 105.2, 1640, 1586, 1508, 1457, 1425, 1377, 1334, 1269, 1242, 1173,
116.7, 127.9, 130.7, 133.0, 139.1, 151.6, 161.0, 168.9 ppm; IR 1143, 1023, 978, 854, 749 cm1; MS (EI, 70 eV): m/z (%): 308.2
(neat): 3277, 2218, 1666, 1619, 1525, 1458, 1438, 1274, 1203, (7) [M]+; HRMS (ESI): m/z [M+H]+ calcd for C18H17N2O3:
1155, 1114, 1067, 1029, 1011, 932, 838 cm1; HRMS (ESI): m/z 309.1234, found: 309.1237. Anal. Calcd for C18H16N2O3: C, 70.12;
[M+H]+ calcd for C18H20N3O3: 326.1499, found: 326.1493; HPLC H, 5.23; N, 9.09. Found: C, 70.02; H, 5.31; N, 8.72.
purity >91%.
4.2.1.54. (E)-2-Cyano-3-[4-(dimethylamino)phenyl]prop-2-ene-
4.2.1.50. (2E,4E)-2-Cyano-4-methyl-5-phenylpenta-2,4-diena- thioamide (73). Starting from 4-(dimethylamino)benzaldehyde
mide (67). Starting from (E)-2-methyl-3-phenylacrylaldehyde (230 mg, 1.54 mmol) and 2-cyanoethanethioamide (165 mg,
(1.46 g, 10.0 mmol) and 2-cyanoacetamide (1.07 mg, 12.7 mmol) 1.65 mmol) compound 73 was obtained after precipitation as a
compound 67 was obtained after precipitation as a colorless solid red solid (265 mg, 74%). 1H NMR (300 MHz, acetone-d6): d = 3.14
(1.33 g, 63%). 1H NMR (300 MHz, acetone-d6): d = 2.39 (d, (s, 6H), 6.85 (m, 2H), 7.97 (m, 2H), 8.32 (br m, 1H), 8.43 (s, 1H),
J = 1.3 Hz, 3H), 7.04 (br m, 2H), 7.31 (bs, 1H), 7.36–7.54 (m, 5H), 8.92 (br m, 1H) ppm; 13C NMR (75 MHz, acetone-d6): d = 40.0,
7.95 (d, J = 1.0 Hz, 1H) ppm; 13C NMR (75 MHz, acetone-d6): 102.9, 112.6, 118.4, 120.1, 134.5, 154.4, 154.6, 194.7 ppm; IR
d = 15.5, 104.6, 117.5, 129.4, 129.7, 130.8, 134.0, 136.7, 146.5, (neat): 3327, 3283, 3150, 2215, 1638, 1605, 1560, 1516, 1409,
157.6, 163.2 ppm; IR (neat): 3464, 3291, 3150, 2206, 1681, 1571, 1365, 1229, 1194, 1169, 1117, 1061, 943, 905, 806 cm1; MS (EI,
1373, 1211, 1111, 1078, 1010, 962, 756 cm1; MS (EI, 70 eV): m/z 70 eV): m/z (%): 231.1 (100) [M]+; HRMS (ESI): m/z [M+H]+ calcd
(%): 212.1 (100) [M]+; HRMS (ESI): m/z [M+H]+ calcd for for C12H14N3S: 232.0903, found: 232.0902. Anal. Calcd for
C13H13N2O: 213.1022, found: 213.1021. Anal. Calcd for C12H13N3S: C, 62.31; H, 5.66; N, 18.17. Found: C, 62.06; H, 5.65;
C13H12N2O: C, 73.56; H, 5.70; N, 13.20. Found: C, 73.45; H, 5.85; N, 18.02.
N, 13.15.
4.2.1.55. (E)-2-Cyano-3-[4-(dimethylamino)phenyl]acrylic acid
4.2.1.51. (2E,4E)-2-Cyano-4-methyl-5-phenyl-N-(tetrahydrofu- (74). Starting from 4-(dimethylamino)benzaldehyde (746 mg,
ran-2-ylmethyl)penta-2,4-dienamide (68). Starting from (E)-2- 5.0 mmol) and cyanoacetic acid (638 mg, 7.5 mmol) compound
methyl-3-phenylacrylaldehyde (390 mg, 2.67 mmol) and 85 74 was obtained after precipitation as an orange solid (600 mg,
(490 mg, 2.91 mmol) compound 68 was obtained after precipita- 56%). 1H NMR (300 MHz, DMSO-d6): d = 3.06 (s, 6H), 6.82 (m,
tion as a colorless solid (300 mg, 38%). 1H NMR (300 MHz, ace- 2H), 7.93 (m, 2H), 8.06 (s, 1H) ppm; 13C NMR (75 MHz, DMSO-
tone-d6): d = 1.62 (m, 1H), 1.80–2.01 (m, 3H), 2.39 (d, J = 1.2 Hz, d6): d = 39.6, 93.4, 111.7, 117.9, 118.4, 133.5, 153.5, 153.8,
3H), 3.35 (m, 1H), 3.49 (m, 1H), 3.68 (m, 1H), 3.83 (m, 1H), 4.02 164.8 ppm; IR (neat): 2215, 1657, 1609, 1561, 1516, 1433, 1384,
(m, 1H), 7.23 (br m, 2H), 7.31 (bs, 1H), 7.35–7.54 (m, 5H), 7.96 1330, 1284, 1248, 1166, 1090, 1063, 941, 812, 709 cm1; HRMS
(m, 1H) ppm; 13C NMR (75 MHz, acetone-d6): d = 15.6, 26.3, 29.4, (ESI): m/z [M+H]+ calcd for C12H13N2O2: 217.0972, found:
44.7, 68.4, 77.9, 104.7, 117.5, 129.4, 129.7, 130.8, 134.0, 136.7, 217.0978; HPLC purity >98%.
146.5, 157.2, 161.6 ppm; IR (neat): 3375, 2977, 2937, 2874, 2209,
1658, 1567, 1523, 1278, 1258, 1207, 1084, 1015, 959, 757 cm1; 4.2.1.56. [4-(Dimethylamino)benzylidene]malononitrile (75).
MS (EI, 70 eV): m/z (%): 296.1 (32) [M]+; HRMS (ESI): m/z [M+H]+ Starting from 4-(dimethylamino)benzaldehyde (373 mg,
calcd for C18H21N2O2: 297.1598, found: 297.1609. Anal. Calcd for 2.50 mmol) and malononitrile (250 mg, 3.78 mmol) compound 75
C18H20N2O2: C, 72.95; H, 6.80; N, 9.45. Found: C, 72.79; H, 6.85; was obtained after precipitation as an orange solid (465 mg,
N, 9.35. 94%). 1H NMR (300 MHz, acetone-d6): d = 3.18 (s, 6H), 6.87 (m,
2H), 7.84 (s, 1H), 7.91 (m, 2H) ppm; 13C NMR (75 MHz, acetone-
4.2.1.52. (2E,4E)-2-Cyano-N-cyclopropyl-5-[4-(dimethylamino)- d6): d = 40.1, 71.2, 112.5, 116.0, 116.7, 120.1, 134.5, 155.5,
phenyl]penta-2,4-dienamide (69). Starting from (E)-3-[4- 159.3 ppm; IR (neat): 2921, 2206, 1607, 1561, 1514, 1385, 1357,
(dimethylamino)phenyl]acrylaldehyde (180 mg, 1.03 mmol) and 1259, 1174, 942, 929, 815 cm1; MS (EI, 70 eV): m/z (%): 197.1
84 (150 mg, 1.21 mmol) compound 69 was obtained after precipi- (97) [M]+; HRMS (ESI): m/z [M+H]+ calcd for C12H12N3: 198.1026,
tation as a red solid (225 mg, 78%). 1H NMR (300 MHz, acetone-d6): found: 198.1027. Anal. Calcd for C12H11N3: C, 73.07; H, 5.62; N,
d = 0.63 (m, 2H), 0.71 (m, 2H), 2.85 (m, 1H), 3.05 (s, 6H), 6.77 (m, 21.30. Found: C, 72.95; H, 5.69; N, 21.13.
7332 C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337
4.2.1.57. (E)-2-Cyano-3-(4-methoxyphenyl)acrylsäure (92). 4.2.2. Genreal procedure for the preparation of compounds 2a–
Starting from anisaldehyde (816 mg, 6.0 mmol) and cyanoacetic 4b, 22a/b
acid (638 mg, 7.50 mmol) compound 92 was obtained after precip- A solution of arylketone (1 equiv), cyanoacetamide (1.0–
itation as a colorless solid (761 mg, 62%). 1H NMR (300 MHz, 1.5 equiv), NH4OAc (0.25 equiv) and acetic acid (0.75 equiv) in tol-
DMSO-d6): d = 3.85 (s, 3H), 7.11 (m, 2H), 8.01 (m, 2H), 8.16 (s, uene (10–25 ml) was refluxed at a Dean-Stark apparatus for 2–5 h.
1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 55.6, 102.4, 114.7, After cooling to room temperature crystallization occurred in the
117.4, 124.4, 132.7, 152.0, 162.6, 164.0 ppm; MS (EI, 70 eV): m/z case of 2a and 4b. The crystals were washed with diethyl ether
(%): 203.0 (100) [M]+. and water. The remaining mixture was evaporated and the residue
was purified by flash chromatography (cyclohexane/ethyl acetate)
4.2.1.58. Methyl N-[(E)-2-cyano-3-(4-methoxyphenyl)prop-2- to isolate the cis and trans isomers.
enoyl]-L-serinate (16). To a solution of 92 (305 mg, 1.50 mmol),
methyl L-serinate (245 mg, 1.58 mmol) and HATU (627 mg, 4.2.2.1. (E)-2-Cyano-3-phenylbut-2-enamide (2a) and (Z)-2-
1.65 mmol) in dry DMF (5 ml) was added DIPEA (0.78 ml, cyano-3-phenylbut-2-enamide (2b). Starting from acetophenone
4.50 mmol) slowly. After 2 h the reaction was quenched with (1.80 g, 15.0 mmol) and 2-cyanoacetamide (1.39 g, 16.5 mmol)
water and the mixture was extracted with ethyl acetate. After compound 2a (470 mg, 17%) and 2b (450 mg, 16%) were obtained
drying over MgSO4 the solvent was evaporated and the residue as colorless solids.
was purified by flash chromatography (cyclohexane/ethyl ace-
tate). Compound 16 was obtained as a colorless solid (288 mg, 4.2.2.1.1. (E)-2-Cyano-3-phenylbut-2-enamide (2a). 1H NMR
63%). 1H NMR (300 MHz, acetone-d6): d = 3.74 (s, 3H), 3.92 (s, (300 MHz, acetone-d6): d = 2.48 (s, 3H), 7.06 (br m, 1H), 7.36 (br
3H), 3.97 (m, 1H), 4.04 (m, 1H), 4.67 (m, 1H), 7.13 (m, 2H), m, 1H), 7.43–7.53 (m, 5H) ppm; 13C NMR (75 MHz, DMSO-d6):
7.42 (br m, 1H), 8.04 (m, 2H), 8.21 (s, 1H) ppm; 13C NMR d = 22.0, 108.5, 116.9, 127.1, 128.6, 129.7, 139.3, 160.8,
(75 MHz, acetone-d6): d = 52.6, 56.1, 56.4, 62.5, 102.1, 115.6, 163.5 ppm; IR (neat): 3371, 3176, 2215, 1655, 1590, 1491, 1429,
117.7, 125.5, 133.8, 152.3, 161.4, 164.4, 171.2 ppm; IR (neat): 1380, 1121, 793, 764, 695 cm1; HRMS (ESI): m/z [M+H]+ calcd
3385, 2954, 2214, 1735, 1645, 1611, 1579, 1514, 1462, 1436, for C11H11N2O: 187.0866, found: 187.0867. Anal. Calcd for
1374, 1314, 1258, 1212, 1177, 1093, 1054, 1024, 958, C11H10N2O: C, 70.95; H, 5.41; N, 15.04. Found: C, 70.90; H, 5.55;
847 cm1; MS (EI, 70 eV): m/z (%): 304.1 (18) [M]+; HRMS N, 14.96.
(ESI): m/z [M+H]+ calcd for C15H17N2O5: 305.1132, found:
305.1133; HPLC purity >98%. 4.2.2.1.2. (Z)-2-Cyano-3-phenylbut-2-enamide (2b). 1H NMR
(300 MHz, acetone-d6): d = 2.44 (s, 3H), 6.78 (br m, 2H), 7.40 (m,
4.2.1.59. N2-[(E)-2-Cyano-3-(4-methoxyphenyl)prop-2-enoyl]- 5H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 24.0, 109.2, 116.7,
N1-cyclopropyl-L-serinamide (17). To a solution of 92 (305 mg, 127.1, 128.3, 129.5, 138.1, 158.7, 163.4 ppm; IR (neat): 3371,
1.50 mmol), 91 (284 mg, 1.58 mmol) and HATU (627 mg, 3176, 2217, 1644, 1591, 1491, 1381, 1279, 1086, 794, 764,
1.65 mmol) in dry DMF (5 ml) was added DIPEA (0.78 ml, 696 cm1; HRMS (ESI): m/z [M+H]+ calcd for C11H11N2O:
4.50 mmol) slowly. After 2 h the reaction was quenched with 187.0866, found: 187.0864; HPLC purity 70% (containing 30% 2a).
water and the mixture was extracted with ethyl acetate. After
drying over MgSO4 the solvent was evaporated and the residue 4.2.2.2. (E)-2-Cyano-4-methyl-3-phenylpent-2-enamide (3a)
was purified by flash chromatography (cyclohexane/ethyl ace- and (Z)-2-cyano-4-methyl-3-phenylpent-2-enamide (3b). Start-
tate). Compound 17 was obtained as a colorless solid (240 mg, ing from isobutyrophenone (5.0 g, 33.7 mmol) and 2-cyanoacet-
48%). 1H NMR (300 MHz, acetone-d6): d = 0.48 (m, 2H), 0.66 amide (2.84 g, 33.8 mmol) compound 3a (40 mg, 0.5%) and 3b
(m, 2H), 2.73 (m, 1H), 3.82 (m, 1H), 3.91 (m, 1H), 3.92 (s, 3H), (425 mg, 6%) were obtained as colorless solids with pure crystals
4.45 (m, 1H), 7.12 (m, 2H), 7.48 (br m, 1H), 8.03 (m, 2H), 8.18 of 3b used for X-ray analysis.
(s, 1H) ppm; 13C NMR (75 MHz, acetone-d6): d = 6.4, 23.3, 56.1,
56.5, 63.0, 102.5, 115.6, 117.8, 125.6, 133.7, 151.9, 161.3, 4.2.2.2.1. (E)-2-Cyano-4-methyl-3-phenylpent-2-enamide (3a).
1
164.3, 171.3 ppm; IR (neat): 3545, 3426, 3266, 3079, 2204, H NMR (300 MHz, acetone-d6): d = 1.02 (d, J = 6.9 Hz, 6H), 3.61
1652, 1590, 1510, 1425, 1254, 1173, 1067, 1015, 835 cm1; MS (sept, J = 6.9 Hz, 1H), 7.07 (br m, 2H), 7.22 (m, 2H), 7.45 (m, 3H)
(EI, 70 eV): m/z (%): 329.2 (6) [M]+; HRMS (ESI): m/z [M+H]+ ppm; 13C NMR (75 MHz, acetone-d6): d = 20.9, 32.8, 110.9, 116.7,
calcd for C17H20N3O4: 330.1448, found: 330.1447; HPLC purity 128.5, 129.0, 129.3, 137.5, 164.0, 171.9 ppm; IR (neat): 3357,
>98%. 3158, 2967, 2223, 1687, 1631, 1581, 1489, 1460, 1440, 1371,
989, 800, 758, 704 cm1; HRMS (ESI): m/z [M+H]+ calcd for
4.2.1.60. 4-[(E)-3-Amino-2-cyano-3-oxoprop-1-en-1-yl]phenyl C13H15N2O: 215.1179, found: 215.1169. Anal. Calcd for
acetate (34). A solution of 21 (188 mg, 1.0 mmol), triethylamine C13H14N2O: C, 72.87; H, 6.59; N, 13.07. Found: C, 72.70; H, 6.59;
(3.0 ml, 21.6 mmol) and acetic anhydride (5.0 ml, 52.9 mmol) in N, 12.99.
acetone (5 ml) and dichloromethane (5 ml) was stirred for
30 min. After quenching with water and evaporation of the solvent 4.2.2.2.2. (Z)-2-Cyano-4-methyl-3-phenylpent-2-enamide (3b).
1
the residue was purified by flash chromatography (cyclohexane/ H NMR (300 MHz, acetone-d6): d = 1.08 (d, J = 6.9 Hz, 6H), 3.32
ethyl acetate). Compound 34 was obtained as a colorless solid (sept, J = 6.9 Hz, 1H), 6.57 (br m, 2H), 7.21 (m, 2H), 7.41 (m, 3H)
(105 mg, 46%). 1H NMR (300 MHz, acetone-d6): d = 2.30 (s, 3H), ppm; 13C NMR (125 MHz, acetone-d6): d = 20.7, 36.5, 111.1 116.3,
7.15 (br m, 2H), 7.35 (m, 2H), 8.07 (m, 2H), 8.23 (s, 1H) ppm; 13C 128.6, 128.8, 129.3, 136.0, 163.6 169.5 ppm; IR (neat): 3454,
NMR (75 MHz, acetone-d6): d = 21.0, 106.4, 117.2, 123.6, 130.6, 3403, 3188, 2973, 2936, 2271, 1672, 1604, 1410, 1378, 1263,
132.6, 151.3, 154.8, 162.6, 169.3 ppm; IR (neat): 3410, 3152, 926, 852, 795, 755, 709 cm1; HRMS (ESI): m/z [M+H]+ calcd for
2218, 1760, 1692, 1595, 1507, 1418, 1380, 1364, 1294, 1199, C13H15N2O: 215.1179, found: 215.1188; HPLC purity >98%.
1166, 1116, 1014, 971, 946, 907, 841, 790 cm1; HRMS (ESI): m/z
[M+H]+ calcd for C12H11N2O3: 231.0764, found: 231.0750. Anal. 4.2.2.3. (E)-2-Cyano-3-phenylhex-2-enamide (4a) and (Z)-2-
Calcd for C12H10N2O3: C, 62.60; H, 4.38; N, 12.17. Found: C, cyano-3-phenylhex-2-enamide (4b). Starting from butyrophe-
62.62; H, 4.53; N, 12.05. none (2.22 g, 15.0 mmol) and 2-cyanoacetamide (1.68 g,
C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337 7333
20.0 mmol) compound 4a (315 mg, 10%) and 4b (295 mg, 9%) were 4.2.3. General procedure for the preparation of compounds 55–
obtained as colorless solids. 60, 63
A solution of fluorobenzaldehyde (1 equiv), cyanoacetamide or
4.2.2.3.1. (E)-2-Cyano-3-phenylhex-2-enamide (4a). 1H NMR derivative (1.05–1.1 equiv) and cyclic amine (2.5–10 equiv) in eth-
(300 MHz, CDCl3): d = 0.91 (t, J = 7.3 Hz, 3H), 1.41 (m, 2H), 3.09 anol (5–10 ml) was reacted in a microwave reactor for 20 min at
(m, 2H), 5.64 (br m, 1H), 6.22 (br m, 1H), 7.36 (m, 2H), 7.45 (m, 160 °C. After cooling to room temperature precipitation occurred
3H) ppm; 13C NMR (75 MHz, CDCl3): d = 14.0, 21.8, 37.0, 105.9, and the precipitate was collected by filtration and washed with
117.7, 127.2, 128.7, 130.1, 139.3, 162.8, 175.4 ppm; IR (neat): ethanol/water (1:1). If precipitation did not occur, the mixture
3410, 3322, 2962, 2220, 2025, 1975, 1697, 1660, 1565, 1440, was evaporated and the residue was purified by flash
1362, 1080, 777, 739, 697 cm1; HRMS (ESI): m/z [M+H]+ calcd chromatography.
for C13H15N2O: 215.1179, found: 215.1180. Anal. Calcd for
C13H14N2O: C, 72.87; H, 6.59; N, 13.07. Found: C, 72.99; H, 6.72; 4.2.3.1. (E)-2-Cyano-3-(4-pyrrolidin-1-ylphenyl)acrylamide (55).
N, 13.01. Starting from 4-fluorobenzaldehyde (372 mg, 3.0 mmol), 2-cyano-
acetamide (265 mg, 3.15 mmol) and pyrrolidine (534 mg,
4.2.2.3.2. (Z)-2-Cyano-3-phenylhex-2-enamide (4b). 1H NMR 7.51 mmol) compound 55 was obtained after precipitation and
(300 MHz, CDCl3): d = 0.94 (t, J = 7.4 Hz, 3H), 1.42 (m, 2H), 2.81 recrystallization from acetone as an orange solid (280 mg, 39%).
1
(m, 2H), 5.37 (br m, 1H), 5.82 (br m, 1H), 7.20 (m, 2H), 7.41 (m, H NMR (300 MHz, acetone-d6): d = 2.05 (m, 5H), 3.42 (m, 4H),
3H) ppm; 13C NMR (75 MHz, CDCl3): d = 13.6, 20.9, 41.5, 108.8, 6.67 (m, 2H), 6.79 (br m, 2H), 7.91 (m, 2H), 8.02 (s, 1H) ppm; 13C
116.1, 126.8, 128.9, 129.8, 136.9, 163.0, 168.7 ppm; IR (neat): NMR (75 MHz, DMSO-d6): d = 24.9, 47.3, 96.4, 111.8, 118.3, 118.4,
3383, 3147, 2962, 2209, 1691, 1593, 1443, 1374, 1292, 1159, 132.8, 150.4, 150.6, 164.0 ppm; IR (neat): 3395, 3151, 2517,
1069, 802, 759, 695 cm1; HRMS (ESI): m/z [M+H]+ calcd for 2159, 2029, 1976, 1660, 1604, 1568, 1523, 1479, 1443, 1396,
C13H15N2O: 215.1179, found: 215.1180. Anal. Calcd for 1344, 1245, 1157, 957, 807 cm1; HRMS (ESI): m/z [M+H]+ calcd
C13H14N2O: C, 72.87; H, 6.59; N, 13.07. Found: C, 73.07; H, 6.69; for C14H16N3O: 242.1288, found: 242.1287; HPLC purity >98%.
N, 13.04.
4.2.3.2. (E)-2-Cyano-3-(4-piperidin-1-ylphenyl)acrylamide (56).
4.2.2.4. (E)-2-Cyano-3-(4-hydroxyphenyl)but-2-enamide (22a) Starting from 4-fluorobenzaldehyde (400 mg, 3.22 mmol), 2-cya-
and (Z)-2-Cyano-3-(4-hydroxyphenyl)but-2-enamide (22b). noacetamide (300 mg, 3.57 mmol) and piperidine (686 mg,
Starting from 4-hydroxyacetophenone (2.04 g, 15.0 mmol) and 2- 8.06 mmol) compound 56 was obtained after precipitation as an
cyanoacetamide (1.39 g, 16.5 mmol) compound 22a (250 mg, 8%) orange solid (350 mg, 43%). 1H NMR (300 MHz, acetone-d6):
was obtained as colorless crystalline solid and 22b (240 mg, 8%) d = 1.66 (m, 6H), 3.48 (m, 4H), 6.86 (br m, 2H), 7.02 (m, 2H), 7.90
as a pale yellow solid. (m, 2H), 8.03 (s, 1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 24.0,
24.9, 47.5, 98.1, 113.4, 118.0, 119.6, 132.7, 150.2, 153.1,
4.2.2.4.1. (E)-2-Cyano-3-(4-hydroxyphenyl)but-2-enamide (22a). 163.8 ppm; IR (neat): 3392, 3174, 2939, 2852, 2220, 1662, 1604,
1
H NMR (300 MHz, acetone-d6): d = 2.46 (s, 3H), 6.92 (m, 2H), 1573, 1518, 1442, 1395, 1354, 1271, 1242, 1197, 1123, 918,
6.94 (br m, 1H), 7.26 (br m, 1H), 7.43 (m, 2H), 8.83 (s, 1H) 805 cm1; HRMS (ESI): m/z [M+H]+ calcd for C15H18N3O:
ppm; 13C NMR (75 MHz, acetone-d6): d = 22.2, 107.2, 116.1, 256.1444, found: 256.1445; HPLC purity >96%.
118.3, 130.1, 132.0, 159.8, 160.0, 164.1 ppm; IR (neat): 3435,
3193, 2224, 1656, 1609, 1587, 1513, 1433, 1370, 1274, 1220, 4.2.3.3. (E)-2-Cyano-3-(4-morpholin-1-ylphenyl)acrylamide (57).
1178, 1144, 1118, 837 cm1; HRMS (ESI): m/z [M+H]+ Starting from 4-fluorobenzaldehyde (621 mg, 5.0 mmol), 2-cyano-
calcd for C11H11N2O2: 203.0815, found: 203.0813; HPLC purity acetamide (441 mg, 5.25 mmol) and morpholine (1.09 g,
>98%. 12.5 mmol) compound 57 was obtained after precipitation as a yel-
low solid (340 mg, 26%). 1H NMR (300 MHz, DMSO-d6): d = 3.33 (m,
4.2.2.4.2. (Z)-2-Cyano-3-(4-hydroxyphenyl)but-2-enamide (22b). 4H), 3.72 (m, 4H), 7.05 (m, 2H), 7.59 (br m, 2H), 7.86 (m, 2H), 8.00
1
H NMR (300 MHz, acetone-d6): d = 2.41 (s, 3H), 6.75 (br m, 2H), (s, 1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 46.5, 65.8, 99.5,
6.85 (m, 2H), 7.30 (m, 2H), 8.79 (bs, 1H) ppm; 13C NMR (75 MHz, 113.5, 117.8, 121.1, 132.4, 150.2, 153.4, 163.6 ppm; IR (neat):
acetone-d6): d = 24.4, 108.8, 116.1, 117.5, 130.1, 131.9, 159.9, 3135, 2208, 1675, 1605, 1569, 1515, 1434, 1361, 1275, 1254,
164.1, 164.9 ppm; IR (neat): 3240, 2224, 1651, 1605, 1566, 1514, 1232, 1193, 1115, 1071, 1031, 928, 811 cm1; HRMS (ESI): m/z
1362, 1268, 1234, 1189, 850, 831 cm1; HRMS (ESI): m/z [M+H]+ [M+H]+ calcd for C14H16N3O2: 258.1237, found: 258.1217; HPLC
calcd for C11H11N2O2: 203.0815, found: 203.0815; HPLC purity purity >98%.
>95% (containing <5% 22a).
4.2.3.4. (E)-2-Cyano-3-(4-piperazin-1-ylphenyl)acrylamide (58).
4.2.2.5. 2-Cyano-2-(2,3-dihydro-1H-inden-1-ylidene)acetamide Starting from 4-fluorobenzaldehyde (372 mg, 3.0 mmol), 2-cyano-
(45). A solution of 1-indanone (397 mg, 3.0 mmol), 2-cyanoacet- acetamide (265 mg, 3.15 mmol) and piperazine (2.58 g,
amide (252 mg, 3.0 mmol), NH4OAc (58 mg, 0.75 mmol) and N- 30.0 mmol) compound 58 was obtained after precipitation as an
methylpiperazine (15 mg, 0.15 mmol) was stirred in ethanol orange solid (320 mg, 42%). 1H NMR (300 MHz, DMSO-d6):
(5 ml) at 60 °C overnight. After the evaporation of the solvent the d = 2.92 (m, 4H), 3.37 (m, 4H), 6.88 (br m, 2H), 7.04 (m, 2H), 7.92
residue was purified by flash chromatography (cyclohexane/ethyl (m, 2H), 8.05 (s, 1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 45.3,
acetate). Compound 45 was obtained as a pale brown solid 47.4, 98.6, 113.4, 117.9, 120.3, 132.5, 150.2, 153.6, 163.7 ppm; IR
(30 mg, 5%). 1H NMR (300 MHz, CDCl3): d = 3.08 (m, 2H), 3.57 (m, (neat): 3382, 3264, 2926, 2213, 1681, 1632, 1605, 1561, 1513,
2H), 5.81 (br m, 1H), 6.34 (br m, 1H), 7–34–7.54 (m, 3H), 8.53 (d, 1434, 1362, 1255, 1233, 1189, 1125, 1017, 966, 910, 814 cm1;
J = 8.2 Hz, 1H) ppm; 13C NMR (75 MHz, CDCl3): d = 30.3, 34.2, HRMS (ESI): m/z [M+H]+ calcd for C14H17N4O: 257.1397, found:
94.7, 110.0, 118.4, 125.8, 127.4, 133.5, 137.3, 153.3, 164.3, 257.1320; HPLC purity >90%.
174.6 ppm; IR (neat): 3450, 3361, 3297, 3157, 2203, 1683, 1613,
1567, 1469, 1429, 1396, 1352, 1307, 1206, 1097, 764 cm1; HRMS 4.2.3.5. (E)-2-Cyano-3-[4-(4-methylpiperazin-1-yl)phenyl]acryl-
(ESI): m/z [M+H]+ calcd for C12H11N2O: 199.0866, found: 199.0867; amide (59). Starting from 4-fluorobenzaldehyde (372 mg,
HPLC purity >93%. 3.0 mmol), 2-cyanoacetamide (265 mg, 3.15 mmol) and N-methyl-
7334 C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337
piperazine (751 mg, 7.5 mmol) compound 59 was obtained after 4.2.4.2. (E)-2-Cyano-N-cyclopropyl-3-(5-morpholin-4-yl-2-
precipitation as an orange solid (300 mg, 37%). 1H NMR furyl)acrylamide (71). Starting from 53 (281 mg, 1.0 mmol) com-
(300 MHz, acetone-d6): d = 2.26 (s, 3H), 2.48 (m, 4H), 3.44 (m, pound 71 was obtained after flash chromatography (cyclohexane/
4H), 6.88 (br m, 2H), 7.05 (m, 2H), 7.92 (m, 2H), 8.05 (s, 1H) ethyl acetate) as a yellow crystalline solid (270 mg, 94%). 1H
ppm; 13C NMR (75 MHz, DMSO-d6): d = 45.6, 46.1, 54.1, 99.0, NMR (300 MHz, DMSO-d6): d = 0.53 (m, 2H), 0.61 (m, 2H), 2.71
113.6, 117.9, 120.5, 132.4, 150.2, 153.2, 163.6 ppm; IR (neat): (m, 1H), 3.44 (m, 4H), 3.71 (m, 4H), 5.77 (d, J = 4.0 Hz, 1H), 7.34
3116, 2807, 2208, 1683, 1607, 1572, 1517, 1448, 1362, 1291, (d, J = 4.0 Hz, 1H), 7.52 (s, 1H), 7.77 (d, J = 4.0 Hz, 1H) ppm; 13C
1254, 1234, 1194, 1145, 1079, 1001, 925, 810, 791 cm1; HRMS NMR (75 MHz, DMSO-d6): d = 5.8, 23.3, 45.6, 65.2, 88.4, 89.7,
(ESI): m/z [M+H]+ calcd for C15H19N4O: 271.1553, found: 118.4, 132.3, 139.6, 162.4, 164.0 ppm; IR (neat): 3345, 3119,
271.1542; HPLC purity >90%. 3023, 2964, 2863, 2188, 1644, 1614, 1542, 1488, 1445, 1408,
1366, 1348, 1319, 1261, 1236, 1173, 1116, 1048, 984, 884, 764,
4.2.3.6. (E)-2-Cyano-N-cyclopropyl-3-[4-(4-methylpiperazin-1- 690, 657 cm1; HRMS (ESI): m/z [M+H]+ calcd for C15H18N3O3:
yl)phenyl]acrylamide (60). Starting from 4-fluorobenzaldehyde 288.1343, found: 288.1346. Anal. Calcd for C15H17N3O3: C, 62.71;
(372 mg, 3.0 mmol), 84 (391 mg, 3.15 mmol) and N-methylpipera- H, 5.96; N, 14.63. Found: C, 62.67; H, 6.11; N, 14.56.
zine (751 mg, 7.5 mmol) compound 60 was obtained after precip-
itation and recrystallization from methanol/water as an orange 4.2.4.3. 3-Morpholin-4-ylbenzaldehyde (97). Starting from 2-
solid (180 mg, 19%). 1H NMR (300 MHz, DMSO-d6): d = 0.56 (m, bromobenzaldehyde (925 mg, 5.0 mmol) compound 97 was ob-
2H), 0.66 (m, 2H), 2.20 (s. 3H), 2.41 (m, 4H), 2.75 (m, 1H), 3.37 tained after flash chromatography (cyclohexane/ethyl acetate) as
(m, 4H), 7.04 (m, 2H), 7.83 (m, 2H), 7.90 (s, 1H), 8.20 (m, 1H) a pale yellow oil (140 mg, 15%). 1H NMR (300 MHz, CDCl3):
ppm; 13C NMR (75 MHz, DMSO-d6): d = 5.7, 23.3, 45.7, 46.2, 54.2, d = 3.20 (m, 4H), 3.85 (m, 4H), 7.14 (m, 1H), 7.37 (m, 3H), 9.93 (s,
99.1, 113.6, 117.6, 120.6, 132.4, 149.6, 153.1, 163.4 ppm; IR (neat): 1H) ppm; 13C NMR (75 MHz, CDCl3): d = 48.7, 66.6, 114.5, 121.5,
3323, 2937, 2842, 2794, 2218, 1665, 1606, 1574, 1509, 1449, 1377, 122.2, 129.7, 137.2, 151.6, 192.6 ppm; MS (EI, 70 eV): m/z (%):
1359, 1277, 1232, 1192, 1008, 920, 819 cm1; HRMS (ESI): m/z 194.1 (86) [M]+.
[M+H]+ calcd for C18H23N4O: 311.1866, found: 311.1854; HPLC
purity >94%. 4.2.4.4. (E)-2-Cyano-3-[4-(piperidin-1-ylcarbonyl)phenyl]acryl-
amide (64). To a solution of 37 (432 mg, 2.0 mmol), HOAt
4.2.3.7. (E)-2-Cyano-3-(2-morpholin-4-ylphenyl)acrylamide (63). (300 mg, 2.20 mmol) and HATU (912 mg, 2.40 mmol) in dry
Starting from 2-fluorobenzaldehyde (372 mg, 3.0 mmol), 2-cyano- DMF (3 ml) was added DIPEA (1.10 ml, 6.0 mmol) slowly at
acetamide (265 mg, 3.15 mmol) and morpholine (653 mg, 0 °C. After 30 min the mixture was allowed to warm up to room
7.5 mmol) compound 63 was obtained after flash chromatography temperature and piperidine (0.19 ml, 2.3 mmol) was added. After
(cyclohexane/ethyl acetate) as a yellow solid (60 mg, 8%). 1H NMR 4 h the reaction was quenched with water and the mixture was
(300 MHz, acetone-d6): d = 2.97 (m, 4H), 3.81 (m, 4H), 7.12 (br m, extracted with ethyl acetate. After drying over MgSO4 the sol-
2H), 7.21 (m, 2H), 7.55 (m, 1H), 8.03 (m, 1H), 8.58 (s, 1H) ppm; vent was evaporated and the residue was purified by flash chro-
13
C NMR (125 MHz, acetone-d6): d = 42.9, 66.8, 110.9, 116.7, matography (ethyl acetate/methanol). Compound 64 was
116.9, 121.2, 125.9, 129.7, 135.5, 143.7, 161.1, 163.2 ppm; IR obtained as a pale yellow solid (345 mg, 61%). 1H NMR
(neat): 3370, 3145, 2211, 1696, 1591, 1481, 1447, 1373, 1346, (300 MHz, DMSO-d6): d = 1.55 (br m, 6H), 3.24 (br m, 2H), 3.58
1298, 1227, 1111, 1068, 937, 768 cm1; MS (EI, 70 eV): m/z (%): (br m, 2H), 7.53 (m, 2H), 7.80 (br m, 1H), 7.94 (br m, 1H),
257.1 (100) [M]+; HRMS (ESI): m/z [M+H]+ calcd for C14H16N3O2: 7.96 (m, 2H), 8.20 (s, 1H) ppm; 13C NMR (75 MHz, DMSO-d6):
258.1237, found: 258.1227; HPLC purity >94%. d = 24.0, 25.2, 25.9, 42.3, 47.9, 107.5, 116.3, 127.3, 130.1, 132.5,
139.9, 149.7, 162.5, 167.7 ppm; IR (neat): 3360, 3160, 2944,
4.2.4. General procedure for the preparation of compounds 70, 2856, 2219, 1694, 1610, 1592, 1440, 1376, 1272, 1206, 1117,
71, 97 1090, 1005, 948, 842 cm1; MS (EI, 70 eV): m/z (%): 283.0 (67)
A solution of arylbromide (1 equiv), morpholine (1.5 equiv), L- [M]+; HRMS (ESI): m/z [M+H]+ calcd for C16H18N3O2: 284.1394,
proline (0.2 equiv), CuI (0.1 equiv) and K2CO3 (2 equiv) in DMSO found: 284.1395; HPLC purity >98%.
(1.5–3 ml) was stirred under a nitrogen atmosphere for 4–20 h at
80 °C. After cooling to room temperature water was added and 4.2.4.5. 3-[(E)-2-Cyano-3-(cyclopropylamino)-3-oxoprop-1-en-
the mixture was extracted with ethyl acetate. After evaporation 1-yl]-N-cyclopropylbenzamide (66). To a solution of 39
of the solvent the residue was purified by flash chromatography. (128 mg, 0.50 mmol), HOAt (75 mg, 0.55 mmol) and HATU
(228 mg, 0.60 mmol) in dry DMF (3 ml) was added DIPEA
4.2.4.1. (E)-2-Cyano-3-(5-morpholin-4-yl-2-thienyl)acrylamide (0.28 ml, 1.5 mmol) slowly at 0 °C. After 30 min the mixture was
(70). Starting from 50 (129 mg, 0.5 mmol) compound 70 was ob- allowed to warm up to room temperature and cyclopropylamine
tained after flash chromatography (cyclohexane/ethyl acetate) as (33 mg, 0.58 mmol) was added. After 4 h the reaction was
an orange crystalline solid (50 mg, 38%). 1H NMR (300 MHz, quenched with water and the mixture was extracted with ethyl
CDCl3): d = 3.39 (m, 4H), 3.84 (m, 4H), 6.14 (d, J = 4.5 Hz, 1H), acetate. After drying over MgSO4 the solvent was evaporated and
7.45 (d, J = 4.5 Hz, 1H), 8.17 (s, 1H) ppm; 13C NMR (75 MHz, the residue was purified by flash chromatography (ethyl acetate/
DMSO-d6): d = 49.3, 65.2, 90.8, 105.2, 118.5, 119.7, 142.7, 144.0, methanol). Compound 66 was obtained as a colorless solid
164.0, 167.0 ppm; IR (neat): 3472, 3103, 2194, 1691, 1573, (75 mg, 51%). 1H NMR (300 MHz, DMSO-d6): d = 0.58 (m, 4H),
1510, 1472, 1442, 1396, 1374, 1357, 1310, 1282, 1243, 1154, 0.70 (m, 4H), 2.78 (m, 1H), 2.85 (m, 1H), 7.63 (t, J = 7.8 Hz, 1H),
1111, 1086, 1027, 934, 893, 849, 762 cm1; MS (EI, 70 eV): m/z 7.95 (m, 1H), 8.07 (m, 1H), 8.13 (s, 1H), 8.28 (m, 1H), 8.55 (m,
(%): 263.0 (100) [M]+; HRMS (ESI): m/z [M+H]+ calcd for 1H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 5.7, 23.1, 23.4, 107.6,
C12H14N3O2S: 264.0801, found: 264.0798. Anal. Calcd for 115.9, 129.1, 129.5, 130.3, 131.4, 132.0, 135.3, 149.4, 162.2,
C12H13N3O2S: C, 54.74; H, 4.98; N, 15.96. Found: C, 54.57; H, 166.5 ppm; IR (neat): 3355, 3293, 2219, 1678, 1633, 1604, 1530,
5.30; N, 15.75. 1360, 1313, 1265, 1227, 1207, 1053, 1023, 965, 849, 683 cm1;
C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337 7335
HRMS (ESI): m/z [M+H]+ calcd for C17H18N3O2: 296.1394, found: 4.2.5.5. (E)-2-Cyano-N-cyclopropyl-5-[4-(dimethylamino)-
296.1392; HPLC purity >95%. phenyl]pent-4-enamide (80). Starting from 69 (141 mg,
0.5 mmol) compound 80 was obtained after precipitation as a pale
4.2.5. General procedure for the preparation of compounds 76– orange solid (110 mg, 78%). 1H NMR (300 MHz, DMSO-d6): d = 0.39
81 (m, 2H), 0.62 (m, 2H), 2.62 (m, 2H), 2.88 (s, 6H), 3.65 (t, J = 7.3 Hz,
To a stirring solution of arylcyanoacrylamide (1 equiv) in meth- 1H), 5.85 (dt, J = 15.7, 7.2 Hz, 1H), 6.37 (d, J = 15.8 Hz, 1H), 6.66 (m,
anol (5–10 ml) was added NaBH4 (5 equiv) at 0 °C. After 1 h the 2H), 7.20 (m, 2H), 8.38 (d, J = 4.0 Hz, 1H) ppm; 13C NMR (75 MHz,
reaction was quenched by the addition of an equivalent volume DMSO-d6): d = 5.5, 5.8, 22.6, 33.5, 37.9, 40.0, 112.2, 118.4, 119.2,
of water. If precipitation occurred, the precipitate was collected 124.4, 127.0, 133.1, 149.9, 165.7 ppm; IR (neat): 3287, 2898,
by filtration and washed with water/methanol (1:1). If precipita- 2247, 2189, 1648, 1611, 1523, 1345, 1292, 1226, 1192, 1166,
tion did not occur, the mixture was evaporated and the residue 1063, 1024, 967, 813 cm1; HRMS (ESI): m/z [M+Na]+ calcd for
was purified by flash chromatography. C17H21N3NaO: 306.1577, found: 306.1570; HPLC purity >98%.
4.2.6.1. 2-Cyano-N-cyclopropylacetamide (84). Starting from Compound 90 was obtained as a colorless solid (896 mg, 36%). 1H
Cyclopropylamine (1.4 ml, 20.0 mmol) and methyl cyanoacetate NMR (300 MHz, CDCl3): d = 0.49 (m, 2H), 0.77 (m, 2H), 1.44 (s,
(1.98, 20.0 mmol) compound 84 was obtained after precipitation 9H), 2.71 (m, 1H), 3.01 (br m, 1H), 3.19 (m, 1H), 3.62 (m, 1H),
as a colorless solid (2.13 g, 86%). 1H NMR (300 MHz, CDCl3): 4.06 (m, 1H), 5.55 (br m, 1H), 6.78 (br m, 1H) ppm; 13C NMR
d = 0.58 (m, 2H), 0.82 (m, 2H), 2.72 (m, 1H), 3.33 (s, 2H), 6.33 (br (75 MHz, CDCl3): d = 6.4, 6.5, 22.5, 28.2, 54.8, 62.8, 80.5, 156.2,
m, 1H) ppm; 13C NMR (75 MHz, acetone-d6): d = 6.2, 23.6, 25.9, 172.8 ppm; MS (EI, 70 eV): m/z (%): 224.2 (1) [M]+.
115.9, 163.8 ppm; MS (EI, 70 eV): m/z (%): 124.0 (29) [M]+.
4.2.6.8. N1-Cyclopropylserineamide hydrochloride (91). To a
4.2.6.2. 2-Cyano-N-(tetrahydrofuran-2-ylmethyl)acetamide (85). solution of 90 (628 mg, 2.57 mmol) in ethyl acetate (15 ml) was
Starting from 1-(tetrahydrofuran-2-yl)methanamine (3.1 ml, added a 2 N solution of HCl in diethyl ether (5 ml, 10.0 mmol)
30.0 mmol) and methyl cyanoacetate (3.00, 30.3 mmol) compound slowly. After 24 h at 4 °C the precipitate was collected by filtration
85 was obtained after addition of water (5 drops) and precipitation and washed with diethyl ether. Compound 91 was obtained as a
as a pale rose solid (4.55 g, 90%). 1H NMR (300 MHz, acetone-d6): colorless solid (337 mg, 73%). 1H NMR (300 MHz, D2O): d = 0.54
d = 1.54 (m, 1H), 1.79–1.98 (m, 3H), 3.20 (m, 1H), 3.35 (m, 1H), (m, 2H), 0.75 (m, 2H), 2.63 (m, 1H), 3.83–4.04 (m, 3H) ppm; 13C
3.58 (s, 2H), 3.65 (m, 1H), 3.78 (m, 1H), 3.89 (m, 1H), 7.41 (br m, NMR (75 MHz, D2O): d = 5.3, 22.0, 54.4, 60.1, 169.1 ppm; HRMS
1H) ppm; 13C NMR (75 MHz, acetone-d6): d = 25.9, 26.3, 29.3, (ESI): m/z [M+H]+ calcd for C6H13N2O2: 145.0972, found: 145.0978.
44.2, 44.3, 68.3, 78.0, 116.1, 162.8 ppm; MS (EI, 70 eV): m/z (%):
169.0 (2) [M+H]+. 4.2.6.9. 4-(Hydroxymethyl)benzaldehyde (93). A solution of
terephthalaldehyde (7.5 g, 56 mmol) in THF (50 ml) was cooled
4.2.6.3. 2-Cyano-N-(pyridin-2-ylmethyl)acetamide (86). Starting to 0 °C. Sodium borohydride (750 mg, 19.8 mmol) was added and
from 1-pyridin-2-ylmethanamine (1.08 g, 10.0 mmol) and methyl the mixture was stirred at room temperature for 1 h. After evapo-
cyanoacetate (990 mg, 10.0 mmol) compound 86 was obtained ration of the solvent, the residue was taken up in ethyl acetate and
after evaporation and washing procedure as a colorless solid the solution was washed with water and brine and dried over
(1.79 g, quant.). 1H NMR (300 MHz, DMSO-d6): d = 3.74 (s, 2H), MgSO4. Upon removal of the solvent, the residue was purified by
4.38 (d, J = 5.9 Hz, 2H), 7.29 (m, 2H), 7.76 (td, J = 7.7, 1.8 Hz, 1H), flash chromatography (cyclohexane/ethyl acetate) to obtain 93 as
8.50 (m, 1H), 8.81 (br m, 1H) ppm; 13C NMR (75 MHz, DMSO-d6): a white solid (3.85 g, 50%). 1H NMR (300 MHz, acetone-d6):
d = 25.3, 44.6, 116.2, 121.2, 122.3, 136.8, 148.9, 157.6, 162.4 ppm; d = 4.49 (t, J = 5.3 Hz, 1H), 4.74, (d, J = 4.4 Hz, 2H), 7.58 (m, 2H),
MS (MALDI-TOF): m/z: 176.2 [M+H]+. 7.88 (m, 2H), 10.01 (s, 1H) ppm; 13C NMR (75 MHz, acetone-d6):
d = 64.1, 127.5, 130.2, 136.5, 150.4, 192.6 ppm; HRMS (ESI): m/z
4.2.6.4. 3-Morpholin-4-yl-3-oxopropanenitrile (87). Starting [M+H]+ calcd for C8H9O2: 137.0597, found: 137.0612.
from morpholine (3.50 ml, 40.2 mmol) and methyl cyanoacetate
(3.96 g, 40.0 mmol) compound 87 was obtained after addition of 4.2.6.10. 4-Hydroxy-3-methoxy-5-nitrobenzaldehyde (94). To a
diethyl ether and precipitation as a pale brown solid (5.53 g, solution of vanillin (5.0 g, 32.9 mmol) in acetic acid (50 ml) was
90%). 1H NMR (300 MHz, acetone-d6): d = 3.50 (m, 4H), 3.63 (m, added nitric acid (60%, 2.6 ml) slowly at 0 °C. Afterwards the solu-
4H), 3.87 (m, 2H) ppm; 13C NMR (75 MHz, acetone-d6): d = 43.1, tion was warmed up to room temperature and allowed to stir for
47.0, 66.9, 115.8, 162.1 ppm; MS (MALDI-TOF): m/z: 155.1 [M+H]+. another 30 min. The precipitate was filtered off and washed with
cold water, to give compound 94 as a yellow solid (5.40 g, 83%).
1
4.2.6.5. 3-(4-Methylpiperazin-1-yl)-3-oxopropanenitrile (88). H NMR (300 MHz, DMSO-d6): d = 3.95 (s, 3H), 7.61 (d, J = 1.8 Hz,
Starting from N-methylpiperazine (2.0 g, 20.0 mmol) and methyl 1H), 8.08 (d, J = 1.9 Hz, 1H), 9.85 (s, 1H) ppm; 13C NMR (75 MHz,
cyanoacetate (1.98 g, 20.0 mmol) compound 88 was obtained after DMSO-d6): d = 56.8, 112.5, 120.8, 126.8, 137.0, 147.7, 150.0,
precipitation as a pale rose solid (2.69 g, 80%). 1H NMR (300 MHz, 190.4 ppm; HRMS (ESI): m/z [MH] calcd for C8H6NO5:
DMSO-d6): d = 2.16 (s, 3H), 2.21–2.32 (m, 4H), 3.28–3.46 (m, 4H), 196.0251, found: 196.0277.
4.01 (s, 2H) ppm; 13C NMR (75 MHz, DMSO-d6): d = 24.7, 41.5,
45.2, 45.5, 54.0, 54.3, 116.1, 161.4 ppm; MS (EI, 70 eV): m/z (%): 4.2.6.11. 3,4-Dihydroxy-5-nitrobenzaldehyde (95). A solution of
167.1 (64) [M]+. 94 (3.5 g, 17.8 mmol) in hydrobromic acid (48%, 32 ml) was re-
fluxed for 4 h. After cooling to room temperature and addition of
4.2.6.6. 2-Cyano-N,N-diethylacetamide (89). A mixture of methyl water and ice, the precipitate was filtered off and washed with cold
cyanoacetate (1.5 g, 15.1 mmol) and diethylamine (3.0 ml, water, to give compound 95 as a brown solid (1.62 g, 50%). 1H NMR
29.0 mmol) was stirred 1 h at 0 °C and at room temperature over- (300 MHz, DMSO-d6): d = 7.46 (d, J = 1.9 Hz, 1H), 7.95 (d, J = 1.9 Hz,
night. Dichloromethane was added, the precipitate was filtered 1H), 9.79 (s, 1H), 10.52 (br m, 2H) ppm; 13C NMR (75 MHz, DMSO-
and discarded; the organic layer was washed with 1 N hydrochloric d6): d = 115.8, 119.6, 127.0, 137.2, 147.2, 148.3, 190.5 ppm; HRMS
acid and sodium bicarbonate solution and dried over MgSO4. After (ESI): m/z [MH] calcd for C7H4NO5: 182.0059, found: 182.0088.
evaporation of the solvent, compound 89 was obtained as a pale
red oil (630 mg, 30%). 1H NMR (300 MHz, acetone-d6): d = 1.07 (t, 4.2.6.12. 4-(Morpholin-4-ylcarbonyl)benzaldehyde (96). To a
J = 7.1 Hz, 3H), 1.20 (t, J = 7.1 Hz, 3H), 3.35 (m, 4H), 3.81 (s, 2H) solution of 4-formylbenzoic acid (750 mg, 5.0 mmol), HOAt
ppm; 13C NMR (75 MHz, acetone-d6): d = 13.1, 14.2, 25.1, 41.1, (750 mg, 5.50 mmol) and HATU (2.28 g, 6.0 mmol) in dry dichloro-
43.0, 116.1, 162.3 ppm; MS (MALDI-TOF): m/z: 141.1 [M+H]+. methane (15 ml) was added DIPEA (2.70 ml, 15.0 mmol) slowly at
0 °C. After 30 min the mixture was allowed to warm up to room
4.2.6.7. N2-(tert-Butyloxycarbonyl)-N1-cyclopropylserineamide temperature and morpholine (0.50 ml, 5.75 mmol) was added.
(90). To a solution of N-(tert-butoxycarbonyl)-L-serine (2.00 g, After 2 h the solvent of the reaction mixture was evaporated and
9.75 mmol) and HATU (4.08 g, 10.7 mmol) in dry DMF (15 ml) the residue was purified by flash chromatography (cyclohexane/
were added cyclopropylamine (0.71 ml, 10.2 mmol) and DIPEA ethyl acetate) to obtain compound 96 as a pale yellow oil
(5.09 ml, 29.3 mmol) slowly. After 2 h the reaction was quenched (950 mg, 87%). 1H NMR (300 MHz, acetone-d6): d = 3.31–3.79 (m,
with water and the mixture was extracted with ethyl acetate. After 8H), 7.63 (m, 2H), 7.99 (m, 2H), 10.09 (s, 1H) ppm; 13C NMR
drying over MgSO4 the solvent was evaporated and the residue was (75 MHz, acetone-d6): d = 38.7, 67.2, 128.6, 130.3, 138.0, 142.6,
purified by flash chromatography (dichloromethane/methanol). 169.1, 192.5 ppm; MS (EI, 70 eV): m/z (%): 219.1 (64) [M]+.
C. Nitsche et al. / Bioorg. Med. Chem. 19 (2011) 7318–7337 7337
4.2.6.13. 4-(Benzyloxy)-3-methoxybenzaldehyde (98). A solution H. Bioorg. Med. Chem. Lett. 2006, 16, 36; (b) Erbel, P.; Schiering, N.; D’Arcy, A.;
Renatus, M.; Kroemer, M.; Lim, S. P.; Yin, Z.; Keller, T. H.; Vasudevan, S. G.;
of vanillin (1.52 g, 10.0 mmol), K2CO3 (5 g, 36.2 mmol) and ben-
Hommel, U. Nat. Struct. Mol. Biol. 2006, 13, 372; (c) Schüller, A.; Yin, Z.; Brian
zylbromide (2.5 ml, 21.0 mmol) in acetone (20 ml) was refluxed Chia, C. S.; Doan, D. N. P.; Kim, H.-K.; Shang, L.; Loh, T. P.; Hill, J.; Vasudevan, S.
for 1 h. After cooling to room temperature and addition of water G. Antivir. Res. 2011, 92, 96.
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a pale yellow solid (2.14 g, 88%). 1H NMR (300 MHz, CDCl3): 6. (a) Villhauer, E. B.; Brinkman, J. A.; Naderi, G. B.; Burkey, B. F.; Dunning, B. E.;
d = 3.94 (s, 3H), 5.24 (s, 2H), 6.98 (d, J = 8.2 Hz, 1H), 7.78–7.46 (m, Prasad, K.; Mangold, B. L.; Russell, M. E.; Hughes, T. E. J. Med. Chem. 2003, 46,
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7H), 9.83 (s, 1H) ppm; 13C NMR (75 MHz, CDCl3): d = 56.1, 70.9, 7. Steuer, C.; Gege, C.; Fischl, W.; Heinonen, K. H.; Bartenschlager, R.; Klein, C. D.
109.3, 112.4, 126.6, 127.2, 128.2, 128.7, 130.3, 136.0, 150.1, Bioorg. Med. Chem. 2011, 19, 4067.
153.6, 190.9 ppm; MS (MALDI-TOF): m/z: 243.1 [M+H]+. 8. (a) Tanaka, Y.; Niwa, S.; Nishioka, H.; Yamanaka, T.; Torizuka, M.; Yoshinaga, K.;
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9. Wang, K.; Nguyen, K.; Huang, Y.; Dömling, A. J. Comb. Chem. 2009, 11, 920.
10. CCDC-#848916 (3b) and #848917 (14) contain the supplementary
Christoph Nitsche acknowledges funding by a fellowship of the crystallographic data for this article. These data can be obtained free of
Studienstiftung des deutschen Volkes. Christian Steuer acknowl- charge from The Cambridge Crystallographic Data Centre via
edges funding by a fellowship of the Konrad Adenauer Stiftung. www.ccdc.cam.ac.uk/data_request/cif.
11. Xu, H.; Yu, X. H.; Sun, L. Y.; Liu, J.; Fan, W.; Shen, Y. J.; Wang, W. Tetrahedron
We thank Franziska Schenk and Florian Mulks for assisting in the
Lett. 2008, 49, 4687.
synthesis of some compounds. We thank Dr. Frank Rominger 12. Zhang, H.; Cai, Q.; Ma, D. W. J. Org. Chem. 2005, 70, 5164.
(Chemistry Department, University of Heidelberg) for performing 13. Steuer, C.; Heinonen, K. H.; Kattner, L.; Klein, C. D. J. Biomol. Screen. 2009, 14,
the small-molecule X-ray analysis. 1102.
14. Tomlinson, S. M.; Watowich, S. J. Antivir. Res. 2011, 89, 127.
15. Bodenreider, C.; Beer, D.; Keller, T. H.; Sonntag, S.; Wen, D. Y.; Yap, L. J.; Yau, Y.
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